Dermal Delivery Compositions Comprising Active Agent-Calcium Phosphate Particle Complexes and Methods of Using the Same

ABSTRACT

Dermal delivery compositions are provided. Aspects of the dermal delivery compositions include the presence of active agent-calcium phosphate particle complexes, where these complexes include uniform, rigid, spherical nanoporous calcium phosphate particles associated with one or more active agents. Also provided are methods of using the compositions in active agent delivery applications.

CROSS-REFERENCE To RELATED APPLICATIONS

Pursuant to 35 U.S.C. §119 (e), this application claims priority to thefiling date of U.S. Provisional Patent Application Ser. No. 61/176,057filed May 6, 2009; the disclosure of which application is hereinincorporated by reference.

INTRODUCTION

A variety of different active agents have been and continue to bedeveloped for use in the treatment of a variety of different conditions,including both disease and non-disease conditions. For suchapplications, an effective amount of the active agent must be deliveredto the subject in need thereof. A variety of different deliveryformulations and routes have been developed, where such routes may varydepending on the nature of the active agent. Typically, less invasivedelivery routes are better tolerated and therefore are more desirable.

One type of delivery route that is of great interest because of itsminimally invasive nature is dermal delivery. In dermal delivery, anactive agent composition is applied to a skin site to deliver the activeagent to the subject. Many dermal delivery technologies currently in useor under evaluation are not entirely satisfactory. For example, certaindermal delivery technologies may disrupt the integrity of the stratumcorneum (Sc) and/or rely on the presence of permeation enhancers, whichcan cause unwanted damage and/or irritation. In addition, certain dermaldelivery technologies may be polymer- and/or liposome basedtechnologies, neither of which technologies truly delivers through theSc. Furthermore these technologies cannot be applied to large molecularweight bio-actives, etc.

As such, there continues to be a need for the development of new dermaldelivery technologies which overcome one or more of the disadvantagesexperienced with current dermal delivery approaches.

SUMMARY

Dermal delivery compositions are provided. Aspects of the dermaldelivery compositions include the presence of active agent-calciumphosphate particle complexes, where these complexes include uniform,rigid, spherical nanoporous calcium phosphate particles associated withone or more active agents. Also provided are methods of using thecompositions in active agent delivery applications.

BRIEF DESCRIPTION OF THE FIGURES

FIGS. 1A to 2B provide scanning electron microscope images of uniform,rigid, spherical, nanoporous calcium phosphate particles that find usein delivery compositions of the invention.

FIG. 3 provides a graphical representation of the particle sizedistribution of uniform, rigid, spherical, nanoporous calcium phosphateparticles that find use in delivery compositions of the invention.

FIG. 4A shows a visual image of active agent attached to calciumphosphate particles. FIG. 4B shows a visual image of active agentattached to calcium phosphate particles.

FIGS. 5A and 5B show the tape strip images following application of a10% calcium phosphate particle slurry to the forearm. FIG. 5A showscalcium phosphate particle penetration to the first layer of the stratumcorneum. FIG. 5B shows calcium phosphate particle penetration to thethird layer of the stratum corneum.

FIG. 6 is an image of mouse skin prior to application of calciumphosphate particles. No Ca⁺⁺ is evident in the upper stratum corneum.

FIG. 7A shows calcium particles in the upper stratum corneum as well assmaller particles in the lower stratum corneum. FIG. 7B shows the lossof integrity of the spherical calcium particles

FIG. 8 is a picture showing penetration of CTC fluorescent throughoutthe stratum corneum following topical application.

FIG. 9 shows the results of STAY-C50 with and without calcium phosphateparticle tape stripping.

FIG. 10 is a table showing amount and percentage of lysozyme with andwithout calcium phosphate particles as measured by tape stripping.

FIG. 11 shows Franz cell transdermal delivery of a riboflavinmonophosphate active agent with and without calcium phosphate particles.

DETAILED DESCRIPTION

Dermal delivery compositions are provided. Aspects of the dermaldelivery compositions include the presence of active agent-calciumphosphate particle complexes, where these complexes include uniform,rigid, spherical nanoporous calcium phosphate particles associated withone or more active agents. Also provided are methods of using thecompositions in active agent delivery applications.

Before the present invention is further described, it is to beunderstood that this invention is not limited to particular embodimentsdescribed, as such may vary. It is also to be understood that theterminology used herein is for the purpose of describing particularembodiments only, and is not intended to be limiting, since the scope ofthe present invention will be limited only by the appended claims.

Where a range of values is provided, it is understood that eachintervening value, to the tenth of the unit of the lower limit unlessthe context clearly dictates otherwise, between the upper and lowerlimit of that range and any other stated or intervening value in thatstated range, is encompassed within the invention. The upper and lowerlimits of these smaller ranges may independently be included in thesmaller ranges and are also encompassed within the invention, subject toany specifically excluded limit in the stated range. Where the statedrange includes one or both of the limits, ranges excluding either orboth of those included limits are also included in the invention.

Certain ranges are presented herein with numerical values being precededby the term “about.” The term “about” is used herein to provide literalsupport for the exact number that it precedes, as well as a number thatis near to or approximately the number that the term precedes. Indetermining whether a number is near to or approximately a specificallyrecited number, the near or approximating unrecited number may be anumber which, in the context in which it is presented, provides thesubstantial equivalent of the specifically recited number.

Methods recited herein may be carried out in any order of the recitedevents which is logically possible, as well as the recited order ofevents.

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which this invention belongs. Although any methods andmaterials similar or equivalent to those described herein can also beused in the practice or testing of the present invention, the preferredmethods and materials are now described.

All publications mentioned herein are incorporated herein by referenceto disclose and describe the methods and/or materials in connection withwhich the publications are cited. The publications discussed herein areprovided solely for their disclosure prior to the filing date of thepresent application. Nothing herein is to be construed as an admissionthat the present invention is not entitled to antedate such publicationby virtue of prior invention. Further, the dates of publication providedmay be different from the actual publication dates which may need to beindependently confirmed.

It must be noted that as used herein and in the appended claims, thesingular forms “a”, “an”, and “the” include plural referents unless thecontext clearly dictates otherwise. It is further noted that the claimsmay be drafted to exclude any optional element. As such, this statementis intended to serve as antecedent basis for use of such exclusiveterminology as “solely,” “only” and the like in connection with therecitation of claim elements, or use of a “negative” limitation.

In further describing various aspects of the invention, the activeagent-calcium phosphate particles according to certain embodiments aredescribed in greater detail, followed by a description of embodiments ofdelivery compositions that include the same, as well as methods ofmaking and using the complexes and delivery compositions that includethe same.

Delivery Compositions

As summarized above, active agent delivery compositions are provided.Active agent delivery compositions of the invention include activeagent-calcium phosphate particle complexes present in a deliveryvehicle. The active agent-calcium phosphate particle complex deliveryvehicle components of the delivery compositions are now reviewedseparately in greater detail.

Active Agent-Calcium Phosphate Particle Complexes

Active agent-calcium phosphate particle complexes that are present indelivery compositions of the invention include uniform, rigid,spherical, nanoporous calcium phosphate particles associated with one ormore active agents. As the particles are associated with one or moreactive agents, one or more active agents are bound to the particles insome manner. The active agent(s) may be bound to the particles via anumber of different associative formats, include but not limited to:ionic binding, covalent binding, Van der Waals interactions, hydrogenbinding interactions, normal phase and reverse phase partitioninteractions, etc. As such, the particles may be described as beingloaded with an amount of one or more active agents. By “loaded” is meantthat the particles include an amount of one or more active agents (inother words an amount of a single active agent or two or more differentactive agents) that is bound to the particles. As the active agent isbound to the particles, the active agent does not dissociate from theparticles in any substantial amount when the particles are present inthe delivery composition. Because substantially none of the active agentdissociates from the particles, any amount that does dissociate is 30%or less, such as 20% or less, e.g., 10% or less, including 5% or less byweight of the originally bound amount of active agent. The amount ofactive agent component (which is made up of one or more distinct activeagents) that is bound to the particles may vary depending on theparticular active agent(s) making up the active agent bound particles,and in certain embodiments ranges from 0.01 to 1000 mg/g, such as from0.1 to 750 mg/g and including 1 to 300 mg/g active agent(s)/gramparticle.

The active agent is reversibly associated with the calcium phosphateparticles. By “reversibly associated” is meant that the active agent isreleased from the calcium phosphate particles following delivery to asubject, e.g., following application a delivery composition thatincludes the complexes to a skin site. As reviewed in greater detailbelow, the calcium phosphate particles of the complexes degrade underacidic conditions, such as under conditions of pH 5 or less, e.g., pH4.9 or less, pH 4.7 or less, pH 4.5 or less, pH 4.3 or less. When theparticles degrade, they release their active agent “payload”. TheStratum corneum (SC), the outer most layer of the skin, is made uproughly 20 layers of cells and is roughly 10 μm in thickness. The pH ofthe SC varies depending on its depth. Its outer most layers vary form pH4.3 to 7.0, depending on the site sampled, or the individuals' sex. ThispH rises to around 7.0 near the Stratum granulosum (SG). This rise ismost dramatic in the last few layers of the SC adjoining the SG, as seenbelow. As such, as complexes of the invention penetrate into the SC,they degrade and concomitantly release any active agent associatedtherewith.

The released active agent retains its desired activity despite havingbeen associated with the calcium phosphate particles in a complex.Accordingly, binding and release of the active agent to the calciumphosphate particles results in substantially little, if any, damage tothe active agent. As such, the activity of the active agent is notdiminished to an extent that adversely impacts its utility, where anyreduction in activity caused by the association to the calcium phosphateparticles that may occur with a given active agent is 10% or less, suchas 5% or less and including 1% or less, e.g., as determined by anactivity assay method, e.g., as described in the Experimental Sectionbelow.

In some embodiments, association of the active agent with the calciumphosphate particles in the complexes preserves one or more desirablefeatures of the active agent, such as stability. In other words, thecomplex stabilizes the active agent, as compared to a control that lacksthe calcium phosphate particles.

Uniform, Rigid, Spherical, Nanoporous Calcium Phosphate Particles

The calcium phosphate particles of the active agent-calcium phosphateparticle complexes are uniform, rigid, spherical, nanoporous calciumphosphate particles. By “uniform” is meant that the shape of theparticles does not vary substantially, such that the particles havesubstantially the same spherical shape. By “rigid” is meant that theparticles are hard, such that they are not pliant. The term “spherical”is employed in its conventional sense to mean a round body whose surfaceis at all points substantially equidistant from the center. Of interestare calcium phosphate particles in which the median diameter is 20 μm orless, such as 10 μm or less, including 5 μm or less, where in someinstances the medium diameter is 4 μm or less, such as 3 μm or less,including 2 μm or less. In a given calcium phosphate particulatecomposition, a distribution of diameters may be present, where in someinstances the majority (such as 60% or more, 75% or more, 90% or more,95% or more) of the particles have diameters that range from 0.01 to 20μm, such as from 0.1 to 10 μm, and including from 0.1 to 2 μm. In someinstances, the proportion of the particles that have an average particlediameter of 2 μm or less is 50% or more by number, such as 70% or moreby number, including 90% or more by number.

The particles are nanoporous. By “nanoporous” is meant that theparticles have a porosity of 30% or more, such as 40% or more, including50% or more, where the porosity may range from 30% to 85%, such as from40% to 70%, including from 45% to 55%, as determined using a mercuryintrusion porosimeter porosity determination protocol as described inASTM D 4284-88 “Standard Test Method for Determining Pore VolumeDistribution of Catalysts by Mercury Intrusion Porosimetry”. Porosity isalso described by “pore volume (ml/g)” and in such instances many rangefrom 0.1 ml/g to 2.0 ml/g. In some cases, the particles have a porositysuch that their internal surface area ranges from 10 m²/g to 150 m²/g,such as from 20 m²/g to 100 m²/g, including 30 m²/g to 80 m²/g, asdetermined using a BET gas adsorption surface area determinationprotocol as described in ASTM D3663-03 Standard Test Method for SurfaceArea of Catalysts and Catalyst Carriers. The pore diameter may vary,ranging in certain instances from 2 to 100 nm, such as 5 to 80 nm,including 10 to 60 nm. In addition, the particles may have a tappingdensity ranging from 0.2 g/cm³ to 0.5 g/cm³, such as from 0.25 g/cm³ to0.45 g/cm³, including from 0.3 g/cm³ to 0.4 g/cm³. The tap density canbe measured by using standard ASTM WK13023—New Determination of TapDensity of Metallic Powders by a Constant Volume Measuring Method.

The particles are, in some instances, chemically pure. By chemicallypure is meant that the particles are made up of substantially one typeof calcium phosphate mineral. In some instances, the calcium phosphateparticles are described by the molecular formula Ca₁₀(PO₄)₆(OH)₂.

In some instances, the particles are ceramic particles. By ceramic ismeant that the particles are produced using a method which includes astep of subjecting the particles to high temperature conditions, wheresuch conditions are illustrated below. High temperatures may range from200 to 1000° C., such as 300 to 900° C. and including 300 to 800° C. Insome embodiments, the particles have a compression rupture strengthranging from 20 to 200 MPa, such as from 50 to 150 MPa, and including 75to 90 MPa, as determined using a SHIMADZU MCT-W500 micro-compressiontesting machine particle strength determination protocol with a particlesintered at temperature of 400° C. to 900° C., as described in EuropeanPatent EP1840661.

In some embodiments, the particles are biodegradable, by which is meantthat the particles degrade in some manner, e.g., dissolve, over timeunder physiological conditions. As the particles of these embodimentsare biodegradable under physiological conditions, they at least begin todissolve at a detectable rate under conditions of pH of 5 or less, suchas 4.5 or less, including 4.3 or less. As such, the particles exhibitsolubility under acidic environments of pH 5 or less, such as uponapplication to the skin.

The calcium phosphate particles are non-toxic, e.g., as determined viaUS-FDA 21 CFR Part 58, non-mutagenic, e.g., as determined byMutagenicity Ames Test, and non-irritating, e.g., as determined via SkinSensitization RIPT (Human).

While the uniform, rigid, spherical, nanoporous calcium phosphateparticles of the delivery compositions may vary in a variety ofdifferent parameters, including as reviewed above, in some embodimentsthe particles employed in the delivery compositions are chemically pureparticles that have a mean diameter of 2 μm.

The uniform, rigid, spherical, nanoporous calcium phosphate particles ofthe delivery compositions of the invention may be prepared using anyconvenient protocol. Examples of fabrication protocols of interestinclude, but are not limited to, those described in U.S. Pat. Nos.4,781,904; 5,039,408; 5,082,566; and 5,158,756; the disclosures of whichare herein incorporated by reference. In one protocol of interest, theparticles are manufactured by spray drying a slurry that includes nanocalcium phosphate (e.g., hydroxyapatite) crystals (which may range from2 nm to 100 nm size range) to produce uniform spherical nanoporouscalcium phosphate particles. The resultant particles are then sinteredfor a period of time sufficient to provide mechanically and chemicallystable rigid spheres. In this step, the sintering temperatures may rangefrom 200° C. to 1000° C., such as 300° C. to 900° C. and including 300°C. to 800° C. for a period of time ranging from 1 hour to 10 hours, suchas 2 hours to 8 hours and including 3 hours to 6 hours. Additionaldetails regarding this method of manufacturing the uniform, rigid,spherical, nanoporous calcium phosphate particles are provided in U.S.Provisional Application Ser. No. 61/108,805, the disclosure of which isherein incorporated by reference.

Active Agents

As summarized above, complexes of the invention include an active agentcomponent (made of a single type of active agent or two or moredifferent types of active agents) bound to the uniform, rigid,spherical, nanoporous calcium phosphate particles. The term “activeagent” refers to any compound or mixture of compounds which produces aphysiological result, e.g., a beneficial or useful result, upon contactwith a living organism, e.g., a mammal, such as a human. Active agentsare distinguishable from other components of the delivery compositions,such as carriers, diluents, lubricants, binders, colorants, etc. Theactive agent may be any molecule, as well as binding portion or fragmentthereof, that is capable of modulating a biological process in a livingsubject. In certain embodiments, the active agent may be a substanceused in the diagnosis, treatment, or prevention of a disease or as acomponent of a medication, cosmetic or cosmeceutical.

The active agent is a compound that interacts with or influences orotherwise modulates a target in a living subject. The target may be anumber of different types of naturally occurring structures, wheretargets of interest include both intracellular and extra-cellulartargets. The active agent may include one or more functional groups thatprovide for structural interaction with the intended target. Functionalgroups of interest include, but are not limited to: groups thatparticipate in hydrogen bonding, hydrophobic-hydrophobic interactions,electrostatic interactions. Specific groups of interest include, but arenot limited to amines, amides, sulfhydryls, carbonyls, hydroxyls,carboxyls, etc.

Active agents of interest may include cyclical carbon or heterocyclicstructures and/or aromatic or polyaromatic structures substituted withone or more of the above functional groups. Also of interest as moietiesof active agents are structures found among biomolecules, includingpeptides, saccharides, fatty acids, steroids, purines, pyrimidines,derivatives, structural analogs or combinations thereof. Such compoundsmay be screened to identify those of interest, where a variety ofdifferent screening protocols are known in the art.

The active agents may be derived from a naturally occurring or syntheticcompound that may be obtained from a wide variety of sources, includinglibraries of synthetic or natural compounds. For example, numerous meansare available for random and directed synthesis of a wide variety oforganic compounds and biomolecules, including the preparation ofrandomized oligonucleotides and oligopeptides. Alternatively, librariesof natural compounds in the form of bacterial, fungal, plant and animalextracts are available or readily produced. Additionally, natural orsynthetically produced libraries and compounds are readily modifiedthrough conventional chemical, physical and biochemical means, and maybe used to produce combinatorial libraries. Known pharmacological agentsmay be subjected to directed or random chemical modifications, such asacylation, alkylation, esterification, amidification, etc. to producestructural analogs.

As such, the active agent may be obtained from a library of naturallyoccurring or synthetic molecules, including a library of compoundsproduced through combinatorial means, i.e., a compound diversitycombinatorial library. When obtained from such libraries, the activeagent employed will have demonstrated some desirable activity in anappropriate screening assay for the activity. Combinatorial libraries,as well as methods for producing and screening such libraries, are knownin the art and described in: U.S. Pat. Nos. 5,741,713; 5,734,018;5,731,423; 5,721,099; 5,708,153; 5,698,673; 5,688,997; 5,688,696;5,684,711; 5,641,862; 5,639,603; 5,593,853; 5,574,656; 5,571,698;5,565,324; 5,549,974; 5,545,568; 5,541,061; 5,525,735; 5,463,564;5,440,016; 5,438,119; 5,223,409, the disclosures of which are hereinincorporated by reference.

Active agents of interest include small, medium and large moleculeactive agents. Small molecule active agents are those active agentshaving a molecular weight ranging from 18 to 2500 daltons, such as 1000to 1500 daltons and including 250 to 1000 daltons. Medium moleculeactive agents are those active agents having a molecular weight rangingfrom 2500 to 10,000 daltons, such as 4,000 to 8,000 daltons andincluding 5000 to 7000 daltons. Large molecule active agents are thoseactive agents having a molecular weight of 10,000 daltons or more, suchas 100,000 daltons or more, where in certain instances these largemolecule active agents range from 1 million to 30 million daltons, suchas 5 million to 20 million daltons and including 10 million to 15million daltons.

In certain embodiments, the active agents are present in their saltforms, such that they carry a charge which allows them to bind to theuniform, rigid, spherical, nanoporous calcium phosphate particles of thedelivery compositions in the desired manner.

Active agents of interest include, but are not limited to, those listedin Appendix A of U.S. Application Ser. No. 61/176,057; the disclosure ofwhich is herein incorporated by reference.

Broad categories of active agents of interest include, but are notlimited to: cardiovascular agents; pain-relief agents, e.g., analgesics,anesthetics, anti-inflammatory agents, etc.; nerve-acting agents;chemotherapeutic (e.g., anti-neoplastic) agents; etc. Active agents ofinterest include, but are not limited to:

antibiotics, such as: aminoglycosides, e.g. amikacin, apramycin,arbekacin, bambermycins, butirosin, dibekacin, dihydrostreptomycin,fortimicin, gentamicin, isepamicin, kanamycin, micronomcin, neomycin,netilmicin, paromycin, ribostamycin, sisomicin, spectinomycin,streptomycin, tobramycin, trospectomycin; amphenicols, e.g.azidamfenicol, chloramphenicol, florfenicol, and theimaphenicol;ansamycins, e.g. rifamide, rifampin, rifamycin, rifapentine, rifaximin;b-lactams, e.g. carbacephems, carbapenems, cephalosporins, cehpamycins,monobactams, oxaphems, penicillins; lincosamides, e.g. clinamycin,lincomycin; macrolides, e.g. clarithromycin, dirthromycin, erythromycin,etc.; polypeptides, e.g. amphomycin, bacitracin, capreomycin, etc.;tetracyclines, e.g. apicycline, chlortetracycline, clomocycline,minocycline, etc.; synthetic antibacterial agents, such as2,4-diaminopyrimidines, nitrofurans, quinolones and analogs thereof,sulfonamides, sulfones;

antifungal agents, such as: polyenes, e.g. amphotericin B, candicidin,dermostatin, filipin, fungichromin, hachimycin, hamycin, lucensomycin,mepartricin, natamycin, nystatin, pecilocin, perimycin; syntheticantifungals, such as allylamines, e.g. butenafine, naftifine,terbinafine; imidazoles, e.g. bifonazole, butoconazole, chlordantoin,chlormidazole, etc., thiocarbamates, e.g. tolciclate, triazoles, e.g.fluconazole, itraconazole, terconazole;

anthelmintics, such as: arecoline, aspidin, aspidinol, dichlorophene,embelin, kosin, napthalene, niclosamide, pelletierine, quinacrine,alantolactone, amocarzine, amoscanate, ascaridole, bephenium,bitoscanate, carbon tetrachloride, carvacrol, cyclobendazole,diethylcarbamazine, etc.;

antimalarials, such as: acedapsone, amodiaquin, arteether, artemether,artemisinin, artesunate, atovaquone, bebeerine, berberine, chirata,chlorguanide, chloroquine, chlorprogaunil, cinchona, cinchonidine,cinchonine, cycloguanil, gentiopicrin, halofantrine, hydroxychloroquine,mefloquine hydrochloride, 3-methylarsacetin, pamaquine, plasmocid,primaquine, pyrimethamine, quinacrine, quinidine, quinine, quinocide,quinoline, dibasic sodium arsenate;

antiprotozoan agents, such as: acranil, tinidazole, ipronidazole,ethylstibamine, pentamidine, acetarsone, aminitrozole, anisomycin,nifuratel, tinidazole, benzidazole, suramin;

cardioprotective agents, e.g., Zinecard (dexrazoxane); blood modifiers,including anticoagulants (e.g., coumadin (warfarin sodium), fragmin(dalteparin sodium), heparin, innohep (tinzaparin sodium), lovenox(enoxaparin sodium), orgaran (danaparoid sodium)) antiplatelet agents(e.g., aggrasta (tirofiban hydrochloride), aggrenox (aspirin/extendedrelease dipyridamole), agrylin (anagrelide hydrochloride), ecotrin(acetylsalicylic acid), folan (epoprostenol sodium), halfprin (entericcoated aspirin), integrlilin (eptifibatide), persantine (dipyridamoleUSP), plavix (clopidogrel bisulfate), pletal (cilostazol), reopro(abciximab), ticlid (ticlopidine hydrochloride)), thrombolytic agents(activase (alteplase), retavase (reteplase), streptase (streptokinase));adrenergic blockers, such as cardura (doxazosin mesylate), dibenzyline(phenoxybenzamine hydrochloride), hytrin (terazosin hydrochloride),minipress (prazosin hydrochloride), minizide (prazosinhydrochloride/polythiazide); adrenergic stimulants, such as aldoclor(methyldopa-chlorothiazide), aldomet (methyldopa, methyldopate HCl),aldoril (methyldopa-hydrochlorothiazide), catapres (clonidinehydrochloride USP, clonidine), clorpres (clonidine hydrochloride andchlorthalidone), combipres (clonidine hydrochloride/chlorthalidone),tenex (guanfacine hydrochloride); alpha/bet adrenergic blockers, such ascoreg (carvedilol), normodyne (labetalol hydrochloride); angiotensinconverting enzyme (ACE) inhibitors, such as accupril (quinaprilhydrochloride), aceon (perindopril erbumine), altace (ramipril),captopril, lotensin (benazepril hydrochloride), mavik (trandolapril),monopril (fosinopril sodium tablets), prinivil (lisinopril), univasc(moexipril hydrochloride), vasotec (enalaprilat, enalapril maleate),zestril (lisinopril); angiotensin converting enzyme (ACE) inhibitorswith calcium channel blockers, such as lexxel (enalaprilmaleate-felodipine ER), lotrel (amlodipine and benazeprilhydrochloride), tarka (trandolapril/verapamil hydrochloride ER);angiotensin converting enzyme (ACE) inhibitors with diuretics, such asaccuretic (quinapril HCl/hydroclorothiazide), lotensin (benazeprilhydrochloride and hydrochlorothiazide USP), prinizide(lisinopril-hydrochlorothiazide), uniretic (moexiprilhydrochloride/hydrochlorothiazide), vaseretic (enalaprilmaleate-hydrochlorothiazide), zestoretic (lisinopril andhydrochlorothiazide); angiotensin II receptor antagonists, such asatacand (candesartan cilexetil), avapro (irbesartan), cozaar (losartanpotassium), diovan (valsartan), micardis (telmisartan), teveten(eprosartan mesylate); angiotensin II receptor antagonists withdiuretics, such as avalide (irbesartan-hydrochlorothiazide), diovan(valsartan and hydrochlorothiazide), hyzaar (losartanpotassium-hydrochlorothiazide); antiarrhythmics, such as Group I (e.g.,mexitil (mexiletine hydrochloride, USP), norpace (disopyramidephosphate), procanbid (procainamide hydrochloride), quinaglute(quinidine gluconate), quinidex (quinidine sulfate), quinidine(quinidine gluconate injection, USP), rythmol (propafenonehydrochloride), tambocor (flecainide acetate), tonocard (tocainideHCl)), Group II (e.g., betapace (sotalol HCl), brevibloc (esmololhydrochloride), inderal (propranolol hydrochloride), sectral (acebutololhydrochloride)), Group III (e.g., betapace (sotalol HCl), cordarone(amiodarone hydrochloride), corvert (ibutilide fumarate injection),pacerone (amiodarone HCl), tikosyn (dofetilide)), Group IV (e.g., calan(verapamil hydrochloride), cardizem (diltiazem HCl), as well asadenocard (adenosine), lanoxicaps (digoxin), lanoxin (digoxin));antilipemic acids, including bile acid sequestrants (e.g., colestid(micronized colestipol hydrochloride), welchol (colesevelamhydrochloride)), fibric acid derivatives (e.g., atromid (clofibrate),lopid (gemfibrozal tablets, USP), tricor (fenofibrate capsules)),HMG-CoA reductase inhibitors (e.g., baycol (cerivastatin sodiumtablets), lescol (fluvastatin sodium), lipitor (atorvastatin calcium),mevacor (lovastatin), pravachol (pravastatin sodium), zocor(simvastatin)), Nicotinic Acid (e.g., Niaspan (niacin extended releasetablets)); beta adrenergic blocking agents, e.g., betapace (sotalolHCl), blocadren (timolol maleate), brevibloc (esmolol hydrochloride),cartrol (carteolol hydrochloride), inderal (propranolol hydrochloride),kerlone (betaxolol hydrochloride), nadolol, sectral (acebutololhydrochloride), tenormin (atenolol), toprol (metoprolol succinate),zebeta (bisoprolol fumarate); beta adrenergic blocking agents withdiuretics, e.g., corzide (nadolol and bendroflumethiazide tablets),inderide (propranolol hydrochloride and hydroclorothiazide), tenoretic(atenolol and chlorthalidone), timolide (timololmaleate-hydrochlorothiazide), ziac (bisoprolol fumarate andhydrochloro-thiazide); calcium channel blockers, e.g., adalat(nifedipine), calan (verapamil hydrochloride), cardene (nicardipinehydrochloride), cardizem (diltiazem HCl), covera (verapamilhydrochloride), isoptin (verapamil hydrochloride), nimotop (nimodipine),norvasc (amlodipine besylate), plendil (felodipine), procardia(nifedipine), sular (nisoldipine), tiazac (diltiazem hydrochloride),vascor (bepridil hydrochloride), verelan (verapamil hydrochloride);diuretics, including carbonic anhydrase inhibitors (e.g., daranide(dichlorphenamide)), combination diuretics (e.g., aldactazide(spironolactone with hydrochlorothiazide), dyazide (triamterene andhydrochlorothiazide), maxzide (triamterene and hydrochlorothiazide),moduretic (amiloride HCl-hydrochlorothiazide)), loop diuretics (demadex(torsemide), edecrin (ethacrynic acid, ethacrynate sodium), furosemide),potassium-sparing diuretics (aldactone (spironolactone), dyrenium(triamterene), midamor (amiloride HCl)), thiazides & related diuretics(e.g., diucardin (hydroflumethiazide), diuril (chlorothiazide,chlorothiazide sodium), enduron (methyclothiazide), hydrodiurilhydrochlorothiazide), indapamide, microzide (hydrochlorothiazide) mykrox(metolazone tablets), renese (polythi-azide), thalitone (chlorthalidone,USP), zaroxolyn (metolazone)); inotropic agents, e.g., digitek(digoxin), dobutrex (dobutamine), lanoxicaps (digoxin), lanoxin(digoxin), primacor (milrinone lactate); activase (alteplaserecombinant); adrenaline chloride (epinephrine injection, USP); demser(metyrosine), inversine (mecamylamine HCl), reopro (abciximab), retavase(reteplase), streptase (streptokinase), tnkase (tenecteplase);vasodilators, including coronary vasodilators (e.g., imdur (isosorbidemononitrate), ismo (isosorbide mononitrate), isordil (isosorbidedinitrate), nitrodur (nitroglycerin), nitrolingual (nitroglycerinlingual spray), nitrostat (nitroglycerin tablets, USP), sorbitrate(isosorbide dinitrate)), peripheral vasodilators & combinations (e.g.,corlopam (fenoldopam mesylate), fiolan (epoprostenol sodium), primacor(milrinone lactate)), vasopressors, e.g., aramine (metaraminolbitartrate), epipen (EpiPen 0.3 mg brand of epinephrine auto injector,EpiPen Jr. 0.15 mg brand of epinephrine auto injector), proamatine(midodrine hydrochloride); etc.

psychopharmacological agents, such as (1) central nervous systemdepressants, e.g. general anesthetics (barbiturates, benzodiazepines,steroids, cyclohexanone derivatives, and miscellaneous agents),sedative-hypnotics (benzodiazepines, barbiturates, piperidinediones andtriones, quinazoline derivatives, carbamates, aldehydes and derivatives,amides, acyclic ureides, benzazepines and related drugs, phenothiazines,etc.), central voluntary muscle tone modifying drugs (anticonvulsants,such as hydantoins, barbiturates, oxazolidinediones, succinimides,acylureides, glutarimides, benzodiazepines, secondary and tertiaryalcohols, dibenzazepine derivatives, valproic acid and derivatives, GABAanalogs, etc.), analgesics (morphine and derivatives, oripavinederivatives, morphinan derivatives, phenylpiperidines,2,6-methane-3-benzazocaine derivatives, diphenylpropylamines andisosteres, salicylates, p-aminophenol derivatives, 5-pyrazolonederivatives, arylacetic acid derivatives, fenamates and isosteres, etc.)and antiemetics (anticholinergics, antihistamines, antidopaminergics,etc.), (2) central nervous system stimulants, e.g. analeptics(respiratory stimulants, convulsant stimulants, psychomotor stimulants),narcotic antagonists (morphine derivatives, oripavine derivatives,2,6-methane-3-benzoxacine derivatives, morphinan derivatives)nootropics, (3) psychopharmacologicals, e.g. anxiolytic sedatives(benzodiazepines, propanediol carbamates) antipsychotics (phenothiazinederivatives, thioxanthine derivatives, other tricyclic compounds,butyrophenone derivatives and isosteres, diphenylbutylamine derivatives,substituted benzamides, arylpiperazine derivatives, indole derivatives,etc.), antidepressants (tricyclic compounds, MAO inhibitors, etc.), (4)respiratory tract drugs, e.g. central antitussives (opium alkaloids andtheir derivatives);

pharmacodynamic agents, such as (1) peripheral nervous system drugs,e.g. local anesthetics (ester derivatives, amide derivatives), (2) drugsacting at synaptic or neuroeffector junctional sites, e.g. cholinergicagents, cholinergic blocking agents, neuromuscular blocking agents,adrenergic agents, antiadrenergic agents, (3) smooth muscle activedrugs, e.g. spasmolytics (anticholinergics, musculotropic spasmolytics),vasodilators, smooth muscle stimulants, (4) histamines andantihistamines, e.g. histamine and derivative thereof (betazole),antihistamines (H1-antagonists, H2-antagonists), histamine metabolismdrugs, (5) cardiovascular drugs, e.g. cardiotonics (plant extracts,butenolides, pentadienolids, alkaloids from erythrophleum species,ionophores, -adrenoceptor stimulants, etc), antiarrhythmic drugs,antihypertensive agents, antilipidemic agents (clofibric acidderivatives, nicotinic acid derivatives, hormones and analogs,antibiotics, salicylic acid and derivatives), antivaricose drugs,hemostyptics, (6) blood and hemopoietic system drugs, e.g. antianemiadrugs, blood coagulation drugs (hemostatics, anticoagulants,antithrombotics, thrombolytics, blood proteins and their fractions), (7)gastrointestinal tract drugs, e.g. digestants (stomachics, choleretics),antiulcer drugs, antidiarrheal agents, (8) locally acting drugs;

chemotherapeutic agents, such as (1) anti-infective agents, e.g.ectoparasiticides (chlorinated hydrocarbons, pyrethins, sulfuratedcompounds), anthelmintics, antiprotozoal agents, antimalarial agents,antiamebic agents, antileiscmanial drugs, antitrichomonal agents,antitrypanosomal agents, sulfonamides, antimycobacterial drugs,antiviral chemotherapeutics, etc., and (2) cytostatics, i.e.antineoplastic agents or cytotoxic drugs, such as alkylating agents,e.g. Mechlorethamine hydrochloride (Nitrogen Mustard, Mustargen, HN2),Cyclophosphamide (Cytovan, Endoxana), Ifosfamide (IFEX), Chlorambucil(Leukeran), Melphalan (Phenylalanine Mustard, L-sarcolysin, Alkeran,L-PAM), Busulfan (Myleran), Thiotepa (Triethylenethiophosphoramide),Carmustine (BiCNU, BCNU), Lomustine (CeeNU, CCNU), Streptozocin(Zanosar) and the like; plant alkaloids, e.g. Vincristine (Oncovin),Vinblastine (Velban, Velbe), Paclitaxel (Taxol), and the like;antimetabolites, e.g. Methotrexate (MTX), Mercaptopurine (Purinethol,6-MP), Thioguanine (6-TG), Fluorouracil (5-FU), Cytarabine (Cytosar-U,Ara-C), Azacitidine (Mylosar, 5-AZA) and the like; antibiotics, e.g.Dactinomycin (Actinomycin D, Cosmegen), Doxorubicin (Adriamycin),Daunorubicin (duanomycin, Cerubidine), Idarubicin (Idamycin), Bleomycin(Blenoxane), Picamycin (Mithramycin, Mithracin), Mitomycin (Mutamycin)and the like, and other anticellular proliferative agents, e.g.Hydroxyurea (Hydrea), Procarbazine (Mutalane), Dacarbazine (DTIC-Dome),Cisplatin (Platinol) Carboplatin (Paraplatin), Asparaginase (Elspar)Etoposide (VePesid, VP-16-213), Amsarcrine (AMSA, m-AMSA), Mitotane(Lysodren), Mitoxantrone (Novatrone), and the like.

Drug compounds of interest are also listed in: Goodman & Gilman's, ThePharmacological Basis of Therapeutics (9th Ed) (Goodman et al. eds)(McGraw-Hill) (1996); and 2001 Physician's Desk Reference.

Specific categories and examples of active agents include, but are notlimited to: those appearing the following table:

Therapeutic Pharmacological Category Class Structural Examples (Does notinclude derivatives) Analgesics Opioid Analgesics Includes drugs such asMorphine, Meperidine and Propoxyphene Non-opioid Includes drugs such asSodium Salicylate, Diflunisal, Para-Aminophenol Analgesics Derivatives,Anthranilic Acid Derivatives, and Phenylpropionic Acid DerivativesAnesthetics Antibacterials Beta-lactam, Cephalosporins Beta-lactam,Penicillins Beta-lactam, Other Includes drugs such as LoracarbefMacrolides Quinolones Sulfonamides Tetracyclines Antibacterials, OtherIncludes drugs such as Trimethoprim, Vancomycin, Lincomycin,Clindamycin, Furazolidone, Nitrofurantoin, Linezolid, Bacitracin,Chloramphenicol, Daptomycin, Fosfomycin, Methenamine, Metronidazole,Mupirocin, Rifaximin, Spectinomycin Anticonvulsants Calcium ChannelIncludes drugs such as Nifedipine Modifying Agents Gamma-aminobutyricIncludes drugs such as Clonazepam, Diazepam, and Phenobarbital Acid(GABA) Augmenting Agents Glutamate Reducing Agents Sodium ChannelInhibitors Antidementia Cholinesterase Agents Inhibitors GlutamatePathway Modifiers Antidementia Agents, Includes drugs such as ErgoloidMesylates Other Antidepressants Monoamino Oxidase (Type A) InhibitorsReuptake Inhibitors Antidepressants, Includes drugs such as Bupropion,Maprotiline, Mirtazapine, Trazodone Other Antiemetics AntifungalsIncludes drugs such as Amphotericin B, and Ketoconazole Antigout AgentsAnti- Glucocorticoids See Adrenal Pharmacologic Class forsimilar/related therapies inflammatories Nonsteroidal Anti- SeeNon-opioid Analgesics Pharmacologic Class for similar/related therapiesinflammatory Drugs (NSAIDs) Antimigraine Abortive See AnalgesicsTherapeutic Category for similar/related therapies Agents ProphylacticSee Autonomic Agents and Cardiovascular Agents Therapeutic Categoriesfor similar/related therapies Antimycobacterials AntitubercularsIncludes drugs such as Isoniazid, Pyridoxine and CycloserineAntimycobacterials, Includes drugs such as Clofazimine, Dapsone,Rifabutin Other Antineoplastics Alkylating Agents Includes drugs such asChlorambucil, Thiotepa, Busulfan, Dacarbazine, and CarmustineAntimetabolites Includes drugs such as Methotrexate, Cytarabine, andMercaptopurine Immune Modulators Includes biotech drugs as variousMonoclonal Antibodies, Cytokines, and Vaccines Interferones andInterleukins Molecular Target Includes drugs such as Vaccines, Antisenseand Gene Tharapies Inhibitors Nucleoside Analogues Includes drugs suchas dIdC, and AZT Protective Agents Includes biotech drugs as VaccinesTopoisomerase Inhibitors Antineoplastics, Includes drugs such asCarboplatin, Cisplatin, Oxaliplatin Other Antiparasitics AnthelminticsIncludes drugs such as Mebendazole, Pyrantel Pamoate, Bithionol, andParomomycin Antiprotozoals Includes drugs such as Chloroquine,Pyrimethamine, Metronidazole, Furazolidone, Melarsoprol, Suramin andTetracyclines Pediculicides/ Includes drugs such as Crotamiton, Lindane,Benzyl Benzoate and Sulfur Scabicides Antiparkinson Catechol O- Agentsmethyltransferase (COMT) Inhibitors Dopamine Agonists Includes drugssuch as Levodopa, and Deprenyl Antiparkinson Includes drugs such asBenztropine, Biperidin, Bromocriptine, Agents, Other Diphenhydramine,Procyclidine, Selegiline, Trihexyphenidyl AntipsychoticsNon-phenothiazines Includes drugs such as Chlorprothixene, andThiothixene Non-phenothiazines/ Includes drugs such as Haloperidol,Molindone, and Loxapine Atypicals Phenothiazines Includes drugs such asFluphenazine Antivirals Anti-cytomegalovirus Includes biotech drugs asVaccines (CMV) Agents Antiherpetic Agents Includes biotech drugs asVaccines and Recombinant Proteins Anti-human immunodeficiency virus(HIV) Agents, Fusion Inhibitors Anti-HIV Agents, Non-nucleoside ReverseTranscriptase Inhibitors Anti-HIV Agents, Nucleoside and NucleotideReverse Transcriptase Inhibitors Anti-HIV Agents, Protease InhibitorsAnti-influenza Agents Includes biotech drugs such as Vaccines, Flumist,and Thymidine Kinase Inhibitors Antivirals, Other Includes drugs such asAdefovir and Ribavirin Anxiolytics Antidepressants Anxiolytics, OtherIncludes drugs such as Buspirone and Meprobamate AutonomicParasympatholytics Agents Parasympathomimetics Sympatholytics SeeCardiovascular Agents and Genitourinary Agents Therapeutic Categoriesfor similar/related therapies Sympathomimetics See Cardiovascular AgentsTherapeutic Category for similar/related therapies Bipolar Agents BloodGlucose Antihypoglycemics Regulators Hypoglycemics, Oral Insulins BloodAnticoagulants Includes drugs such as Acetaminophen, CoumarinDerivatives, Aspirin, Products/ Blood Formation Heparin, and IndandioneDerivatives Modifiers/ Products Volume Coagulants Expanders PlateletAggregation Inhibitors Cardiovascular Alpha-adrenergic See AutonomicAgents Therapeutic Category for similar/related therapies AgentsAgonists Alpha-adrenergic Includes drugs such as Phenolamine Mesylate,and Prazosin HCl Blocking Agents Antiarrhythmics Includes drugs such asBretylium, Digitalis, Quinidine, and Atropine Beta-adrenergic Includesdrugs such as Atenolol and related compounds Blocking Agents CalciumChannel Includes drugs such as Nifedipine Blocking Agents Direct CardiacInotropics Diuretics Includes drugs such as Furosemide, andSpironolactone Dyslipidemics Renin-angiotensin- Includes drugs such asCaptopril, and Saralasin Acetate aldosterone System InhibitorsVasodilators Includes drugs such as Sodium Nitroprusside, NitroglycerineCentral Nervous Amphetamines System Agents Non-amphetamines Dental andIncludes such drugs as CHG Oral Agents Dermatological DermatologicalIncludes drugs such as Lidocaine, Dibucaine, and Diperodon AgentsAnesthetics Dermatological Includes drugs such as Bacitracin,Chlorotetracycline, and Erythromycin Antibacterials DermatologicalIncludes drugs such as Haloprogin, Tolnaftate, Imidazoles, and PolyeneAntifungals Antibiotics Dermatological Anti- Includes drugs such asHydrocortisone, Amcinonide, and Desonide inflammatories DermatologicalIncludes drugs such as Benzocaine, Lidocaine, Pramoxine,Diphenhydramine, Antipruritic Agents and Hydrocortisone DermatologicalHIV-Inhibitors of reverse transcriptase (Nucleoside analogs,Non-nucleoside Antivirals analogs, and Nucleotide analogs), Viralpackaging inhibitors (Protease Inhibitors), Fusion Inhibitors, HerpesVirus-Nucleoside analogs (Acyclovir, Valacyclovir, Famciclovir andPenciclovir), Interferone Alpha, and Imiquimod Dermatological Includesdrugs such as Urea, and Salicylic Acid Keratolytics DermatologicalIncludes drugs such as Vinblastine, and Vincristine Mitotic InhibitorsDermatological Includes drugs such as Hydroquinone and TrioxsalenPhotochemotherapy Agents Dermatological Includes drugs such as TretinoinRetinoids Dermatological Tar Includes drugs such as Anthraquinonederivatives (Anthralin) Derivatives Dermatological Includes drugs suchas Calcitriol, and Calcipotriol Vitamin D Analogs DermatologicalIncludes drugs such as Collagenase, Sutilains and Dextranomers WoundCare Agents Dermatological Includes drugs such as Benzoyl Peroxide, andSalicylic Acid Antiacne Dermatological Includes actives such as3_Benzylidene_Camphors, 2-phenylbenzimidazole- UVA/UVB Block 5-sulfonicacid, Octyl Salicylate, Homosalate, Octylmethyl PABA,, OctylMethoxycinnamate, Octocrylene, Oxybenzone, Menthyl Anthranilate,Titanium Dioxide, Zinc Oxide, Avobenzone Deterrents/ Alcohol DeterrentsReplacements Enzyme Replacements/ Modifiers GastrointestinalAntispasmodics, Includes drugs such as Cimetidine, and Ranitidine AgentsGastrointestinal Histamine2 (H2) Blocking Agents Irritable BowelSyndrome Agents Protectants Proton Pump Inhibitors GastrointestinalIncludes drugs such as Sevelamer, Ursodiol, Antisense, Vaccines and MabAgents, Other and their fragments Genitourinary Antispasmodics, AgentsUrinary Benign Prostatic See Autonomic Agents and Cardiovascular AgentsTherapeutic Categories for Hypertrophy Agents similar/related therapiesImpotence Agents Prostaglandins See Hormonal Agents,Stimulant/Replacement/Modifying TherapeuticCategory for similar/relatedtherapies Hormonal Adrenal See Anti-inflammatories Therapeutic Categoryfor similar/related therapies Agents, Parathyroid/Metabolic Stimulant/Bone Disease Agents Replacement/ Pituitary Modifying Prostaglandins SeeGenitourinary Agents Therapeutic Category for similar/related therapiesSex Hormones/Modifiers Thyroid Includes drugs such as LevothyroxineSodium, and Methimazole Hormonal Adrenal Agents, Suppressant PituitaryIncludes biotech drugs as hGH Sex Includes biotech drugs as EstradiolHormones/Modifiers Thyroid Immunological Immune Stimulants Includesbiotech drugs as various Monoclonal Antibodies, Interferones and AgentsInterleukins Immune Includes biotech drugs as various MonoclonalAntibodies, Interferones and Suppressants Interleukins ImmunomodulatorsIncludes biotech drugs as various Monoclonal Antibodies, Interferonesand Interleukins Inflammatory Glucocorticoids See Hormonal Agents,Stimulant/Replacement/Modifying Therapeutic Bowel Disease Category forsimilar/related therapies Agents Salicylates Sulfonamides SeeAntibacterials Therapeutic Category for similar/related therapiesOphthalmic Ophthalmic Anti- Includes drugs such as Cromolyn Agentsallergy Agents Ophthalmic Includes drugs such as Bacitracin,Chloramphenicol, Erythromycin, and Antibacterials Polymyxin B SulfateOphthalmic Includes drugs such as Amphotericin B, Miconazole, Natamycinand Nystatin Antifungals Ophthalmic Includes drugs such as PilocarpineHCl, Carbachol, Physostigmine Salicylate, Antiglaucoma AgentsIsoflurophate, and Acetazolamide Ophthalmic Anti- Includes drugs such asHydrocortisone, Dexamethasone, and Medrysone inflammatories OphthalmicAntivirals Includes drugs such as Idoxuridine, Trifluridine, Antisense,and Vidarabine Ophthalmics, Other Includes drugs such as FormivirsenOtic Agents Otic Antibacterials Includes drugs such as Chloramphenicol,Neomycin Sulfate, and Polymyxins Otic Anti- inflammatories RespiratoryAntihistamines Tract Agents Antileukotrienes Bronchodilators,Anticholinergic Bronchodilators, Anti- Includes drugs such asCorticosteroid derivatives inflammatories Bronchodilators,Phosphodiesterase 2 Inhibitors (Xanthines) Bronchodilators, Includesdrugs such as Albuterol, Terbutaline, and Isoproterenol SympathomimeticMast Cell Stabilizers Includes drugs such as Cromolyn Sodium MucolyticsRespiratory Tract Includes drugs such as Alpha-1-proteinase Inhibitor,Human; Benzonatate; Agents, Other Guaifenesin; Iodinated Glycerol;Potassium Iodide; Tetrahydrozoline Sedatives/ Hypnotics Skeletal MuscleIncludes drugs such as Carisoprodol, Chlorphenesin Carbamate, RelaxantsChlorzoxazone, and Cyclobenzaprine HCl Therapeutic Electrolytes/MineralsNutrients/ Vitamins Minerals/ Electrolytes Toxicologic OpioidAntagonists Agents Erectial Tadalafil, sildenafil, vardenafildysfunction Agents

Specific compounds of interest also include, but are not limited to:

Hydrocodone/Acetaminophen Lipitor Azithromycin Nexium Simvastatin AdvairDiskus Oxycodone ER Prevacid Sertraline Plavix Fentanyl TransdermalSingulair Amlodipine Besylate Seroquel Fexofenadine Effexor XRAmoxicillin/Pot Clav Lexapro Omeprazole Actos Gabapentin ProtonixFluticasone Nasal Vytorin Lisinopril Topamax Oxycodone w/AcetaminophenRisperdal Metoprolol Succinate Abilify Metformin Cymbalta NovoLogLamictal Amlodipine Besylate Zyprexa Levothyroxine Levaquin ZolpidemTartrate Celebrex Amoxicillin Zetia Ondansetron Valtrex ParoxetineCrestor Alprazolam Fosamax Lovastatin Zyrtec Albuterol Aerosol LantusFluoxetine Adderall XR Lorazepam Diovan Warfarin Avandia PravastatinTricor Cefdinir Aciphex Atenolol Diovan HCT HydrochlorothiazideOxyContin Tramadol Concerta Clonazepam Coreg Cephalexin Flomax BupropionSR Lyrica Oxycodone Wellbutrin XL Propoxyphene-N/Acetaminophen AriceptLisinopril/Hydrochlorothiazide Imitrex Oral Finasteride AmbienCitalopram HBr Lotrel Nifedipine ER Nasonex Cyclobenzaprine Toprol XLFurosemide Oral Ambien CR Carisoprodol Enbrel Morphine Sulfate ERSpiriva Ciprofloxacin HCl Viagra Metoprolol Tartrate Lidoderm PrednisoneOral Actonel Cartia XT Chantix Amphetamine Salt Cmb Norvasc ClindamycinSystemic Lovenox Nabumetone Provigil Potassium Chloride LunestaOndansetron ODT Altace Diltiazem CD Keppra Verapamil SR Geodon OralAlbuterol Nebulizer Solution Cozaar Felodipine ER Detrol LA QuinaprilAtripla Clopidogrel Truvada Ibuprofen CellCept Ranitidine HCl PulmicortRespules Glyburide/Metformin HCl Humalog Minocycline Depakote ERTriamterene w/Hydrochlorothiazide Depakote Enalapril Premarin TabsOxybutynin Chl ER Synthroid Tramadol HCl/Acetaminophen Niaspan MeloxicamByetta Acetaminophen w/Codeine Budeprion XL Spironolactone StratteraHydroxyzine Combivent Naproxen Trileptal Glipizide ER Yasmin 28Trazodone HCl Flovent HFA Fluconazole Skelaxin Mirtazapine PrografPromethazine Tabs Arimidex Phentermine Evista Glyburide HyzaarTizanidine HCl Namenda Diazepam Januvia Venlafaxine Humira Metformin HClER Cialis Buspirone HCl Reyataz Diclofenac Sodium Xalatan DoxycyclineOmnicef Gemfibrozil Avelox Cefprozil ProAir HFA Propranolol HCl AsacolPhenytoin Sodium Ext Benicar HCT Isosorbide Mononitrate Fentanyl OralCitra Clarithromycin Requip Clozapine Boniva Vancocin HCl CaduetGlimepiride Avapro Clotrimazole/Betamethasone Gleevec Carbidopa/LevodopaKaletra Mupirocin Ortho TriCyclen Lo Desmopressin Acetate BenicarNitrofurantoin Monohydrate AndroGel Clonidine Xopenex Clarithromycin ERProcrit Trimethoprim Sulfate Lamisil Oral Nifedical XL AvalideCarvedilol Nasacort AQ Methotrexate Combivir Hydrocodone/IbuprofenAllegra-D 12 Hour Methylprednisolone Tabs Duragesic Etodolac CopaxoneNifedipine RenaGel Bupropion ER Femara Nystatin Systemic EnbrelSureclick Benazepril NovoLog Mix 70/30 Zegerid Clarinex CefuroximeAxetil Aldara Amitriptyline Forteo Bupropion XL Suboxone ClobetasolAvodart Acyclovir Paxil CR Benzonatate Norvir Allopurinol AvandametPenicillin VK Restasis Temazepam Avonex Baclofen Sensipar TretinoinTarceva Sulfamethoxazole/Tri Patanol Terbinafine HCL Yaz Methadone HClNon-Injection Lovaza Amiodarone Mirapex Ketoconazole Topical Focalin XRHydroxy-chloroquine Cosopt Nitrofurantoin Macrocrystals ZyvoxTriamcinolone Acetonide Top Epzicom Lithium Carbonate NuvaRing TerazosinActiq Itraconazole Foxamax Plus D Hydralazine Actoplus MetButalbital/Acetaminophen/Caffeine Lumigan Labetalol Rhinocort AquaFosinopril Sodium Solodyn Cilostazol Thalomid Mometasone Topical FuzeonDoxazosin Astelin Clindamycin Topical BenzaClin Metoclopramide RelpaxMedroxyprogesterone Injection Viread Megestrol Oral Suspension CasodexFolic Acid Vigamox Zostavax Vesicare Nitroglycerin Humalog Mix 75/25 PnBisoprolol/Hydrochlorothiazide Trizivir Polyethylene Glycol Budeprion SRPrednisolone Sd Phosphate Oral Xeloda Azathioprine Sustiva CalcitriolLevitra Torsemide Endocet Glipizide Risperdal Consta Sotalol AggrenoxZonisamide Humira Pen Hydromorphone HCl Kadian Potassium ChlorideDifferin Oxcarbazepine Catapres-TTS Diltiazem SR Alphagan P AlbuterolSulfate/Ipratropium Tussionex Metronidazole Tabs Zyrtec SyrupCabergoline Maxalt Cyclosporine Zoloft Estradiol Oral PrilosecMethocarbamol Ciprodex Otic Tamoxifen Temodar Promethazine/CodeineTobraDex Ursodiol Zyrtec-D Mercaptopurine Welchol Ribavirin Maxalt MLTFamotidine Asmanex PhosLo Atacand Indomethacin SR Coumadin TabsLamotrigine Dovonex Cefadroxil Klor-Con Ipratropium Br NebulizerSolution Pegasys Fluvoxamine Ultram ER Methylphenidate BetaseronMetolazone Zovirax Topical Microgestin Fe 1/20 Trinessa DexamphetamineSulfate Pulmozyme Diltiazem ER Neupogen Clindesse Humulin N FlecainideAcetate Micardis HCT Metronidazole Top Ortho Evra Microgestin Fe 1/20Allegra-D 12 Hours Evoclin Fentora Primidone Enablex Fluocinonide FamvirTerconazole Avinza Carbidopa/Levodopa ER Prempro Leflunomide Coreg CRMidodrine HCl Marinol

Specific compounds of interest also include, but are not limited to:

antineoplastic agents, as disclosed in U.S. Pat. Nos. 5,880,161,5,877,206, 5,786,344, 5,760,041, 5,753,668, 5,698,529, 5,684,004,5,665,715, 5,654,484, 5,624,924, 5,618,813, 5,610,292, 5,597,831,5,530,026, 5,525,633, 5,525,606, 5,512,678, 5,508,277, 5,463,181,5,409,893, 5,358,952, 5,318,965, 5,223,503, 5,214,068, 5,196,424,5,109,024, 5,106,996, 5,101,072, 5,077,404, 5,071,848, 5,066,493,5,019,390, 4,996,229, 4,996,206, 4,970,318, 4,968,800, 4,962,114,4,927,828, 4,892,887, 4,889,859, 4,886,790, 4,882,334, 4,882,333,4,871,746, 4,863,955, 4,849,563, 4,845,216, 4,833,145, 4,824,955,4,785,085, 476,925, 4,684,747, 4,618,685, 4,611,066, 4,550,187,4,550,186, 4,544,501, 4,541,956, 4,532,327, 4,490,540, 4,399,283,4,391,982, 4,383,994, 4,294,763, 4,283,394, 4,246,411, 4,214,089,4,150,231, 4,147,798, 4,056,673, 4,029,661, 4,012,448;

psychopharmacological/psychotropic agents, as disclosed in U.S. Pat.Nos. 5,192,799, 5,036,070, 4,778,800, 4,753,951, 4,590,180, 4,690,930,4,645,773, 4,427,694, 4,424,202, 4,440,781, 5,686,482, 5,478,828,5,461,062, 5,387,593, 5,387,586, 5,256,664, 5,192,799, 5,120,733,5,036,070, 4,977,167, 4,904,663, 4,788,188, 4,778,800, 4,753,951,4,690,930, 4,645,773, 4,631,285, 4,617,314, 4,613,600, 4,590,180,4,560,684, 4,548,938, 4,529,727, 4,459,306, 4,443,451, 4,440,781,4,427,694, 4,424,202, 4,397,853, 4,358,451, 4,324,787, 4,314,081,4,313,896, 4,294,828, 4,277,476, 4,267,328, 4,264,499, 4,231,930,4,194,009, 4,188,388, 4,148,796, 4,128,717, 4,062,858, 4,031,226,4,020,072, 4,018,895, 4,018,779, 4,013,672, 3,994,898, 3,968,125,3,939,152, 3,928,356, 3,880,834, 3,668,210;

cardiovascular agents, as disclosed in U.S. Pat. Nos. 4,966,967,5,661,129, 5,552,411, 5,332,737, 5,389,675, 5,198,449, 5,079,247,4,966,967, 4,874,760, 4,954,526, 5,051,423, 4,888,335, 4,853,391,4,906,634, 4,775,757, 4,727,072, 4,542,160, 4,522,949, 4,524,151,4,525,479, 4,474,804, 4,520,026, 4,520,026, 5,869,478, 5,859,239,5,837,702, 5,807,889, 5,731,322, 5,726,171, 5,723,457, 5,705,523,5,696,111, 5,691,332, 5,679,672, 5,661,129, 5,654,294, 5,646,276,5,637,586, 5,631,251, 5,612,370, 5,612,323, 5,574,037, 5,563,170,5,552,411, 5,552,397, 5,547,966, 5,482,925, 5,457,118, 5,414,017,5,414,013, 5,401,758, 5,393,771, 5,362,902, 5,332,737, 5,310,731,5,260,444, 5,223,516, 5,217,958, 5,208,245, 5,202,330, 5,198,449,5,189,036, 5,185,362, 5,140,031, 5,128,349, 5,116,861, 5,079,247,5,070,099, 5,061,813, 5,055,466, 5,051,423, 5,036,065, 5,026,712,5,011,931, 5,006,542, 4,981,843, 4,977,144, 4,971,984, 4,966,967,4,959,383, 4,954,526, 4,952,692, 4,939,137, 4,906,634, 4,889,866,4,888,335, 4,883,872, 4,883,811, 4,847,379, 4,835,157, 4,824,831,4,780,538, 4,775,757, 4,774,239, 4,771,047, 4,769,371, 4,767,756,4,762,837, 4,753,946, 4,752,616, 4,749,715, 4,738,978, 4,735,962,4,734,426, 4,734,425, 4,734,424, 4,730,052, 4,727,072, 4,721,796,4,707,550, 4,704,382, 4,703,120, 4,681,970, 4,681,882, 4,670,560,4,670,453, 4,668,787, 4,663,337, 4,663,336, 4,661,506, 4,656,267,4,656,185, 4,654,357, 4,654,356, 4,654,355, 4,654,335, 4,652,578,4,652,576, 4,650,874, 4,650,797, 4,649,139, 4,647,585, 4,647,573,4,647,565, 4,647,561, 4,645,836, 4,639,461, 4,638,012, 4,638,011,4,632,931, 4,631,283, 4,628,095, 4,626,548, 4,614,825, 4,611,007,4,611,006, 4,611,005, 4,609,671, 4,608,386, 4,607,049, 4,607,048,4,595,692, 4,593,042, 4,593,029, 4,591,603, 4,588,743, 4,588,742,4,588,741, 4,582,854, 4,575,512, 4,568,762, 4,560,698, 4,556,739,4,556,675, 4,555,571, 4,555,570, 4,555,523, 4,550,120, 4,542,160,4,542,157, 4,542,156, 4,542,155, 4,542,151, 4,537,981, 4,537,904,4,536,514, 4,536,513, 4,533,673, 4,526,901, 4,526,900, 4,525,479,4,524,151, 4,522,949, 4,521,539, 4,520,026, 4,517,188, 4,482,562,4,474,804, 4,474,803, 4,472,411, 4,466,979, 4,463,015, 4,456,617,4,456,616, 4,456,615, 4,418,076, 4,416,896, 4,252,815, 4,220,594,4,190,587, 4,177,280, 4,164,586, 4,151,297, 4,145,443, 4,143,054,4,123,550, 4,083,968, 4,076,834, 4,064,259, 4,064,258, 4,064,257,4,058,620, 4,001,421, 3,993,639, 3,991,057, 3,982,010, 3,980,652,3,968,117, 3,959,296, 3,951,950, 3,933,834, 3,925,369, 3,923,818,3,898,210, 3,897,442, 3,897,441, 3,886,157, 3,883,540, 3,873,715,3,867,383, 3,873,715, 3,867,383, 3,691,216, 3,624,126;

antimicrobial agents as disclosed in U.S. Pat. Nos. 5,902,594,5,874,476, 5,874,436, 5,859,027, 5,856,320, 5,854,242, 5,811,091,5,786,350, 5,783,177, 5,773,469, 5,762,919, 5,753,715, 5,741,526,5,709,870, 5,707,990, 5,696,117, 5,684,042, 5,683,709, 5,656,591,5,643,971, 5,643,950, 5,610,196, 5,608,056, 5,604,262, 5,595,742,5,576,341, 5,554,373, 5,541,233, 5,534,546, 5,534,508, 5,514,715,5,508,417, 5,464,832, 5,428,073, 5,428,016, 5,424,396, 5,399,553,5,391,544, 5,385,902, 5,359,066, 5,356,803, 5,354,862, 5,346,913,5,302,592, 5,288,693, 5,266,567, 5,254,685, 5,252,745, 5,209,930,5,196,441, 5,190,961, 5,175,160, 5,157,051, 5,096,700, 5,093,342,5,089,251, 5,073,570, 5,061,702, 5,037,809, 5,036,077, 5,010,109,4,970,226, 4,916,156, 4,888,434, 4,870,093, 4,855,318, 4,784,991,4,746,504, 4,686,221, 4,599,228, 4,552,882, 4,492,700, 4,489,098,4,489,085, 4,487,776, 4,479,953, 4,477,448, 4,474,807, 4,470,994,4,370,484, 4,337,199, 4,311,709, 4,308,283, 4,304,910, 4,260,634,4,233,311, 4,215,131, 4,166,122, 4,141,981, 4,130,664, 4,089,977,4,089,900, 4,069,341, 4,055,655, 4,049,665, 4,044,139, 4,002,775,3,991,201, 3,966,968, 3,954,868, 3,936,393, 3,917,476, 3,915,889,3,867,548, 3,865,748, 3,867,548, 3,865,748, 3,783,160, 3,764,676,3,764,677;

anti-inflammatory agents as disclosed in U.S. Pat. Nos. 5,872,109,5,837,735, 5,827,837, 5,821,250, 5,814,648, 5,780,026, 5,776,946,5,760,002, 5,750,543, 5,741,798, 5,739,279, 5,733,939, 5,723,481,5,716,967, 5,688,949, 5,686,488, 5,686,471, 5,686,434, 5,684,204,5,684,041, 5,684,031, 5,684,002, 5,677,318, 5,674,891, 5,672,6205,665,752, 5,656,661, 5,635,516, 5,631,283, 5,622,948, 5,618,835,5,607,959, 5,593,980, 5,593,960, 5,580,888, 5,552,424, 5,552,4225,516,764, 5,510,361, 5,508,026, 5,500,417, 5,498,405, 5,494,927,5,476,876, 5,472,973, 5,470,885, 5,470,842, 5,464,856, 5,464,849,5,462,952, 5,459,151, 5,451,686, 5,444,043, 5,436,265, 5,432,181,RE034918, 5,393,756, 5,380,738, 5,376,670, 5,360,811, 5,354,768,5,348,957, 5,347,029, 5,340,815, 5,338,753, 5,324,648, 5,319,099,5,318,971, 5,312,821, 5,302,597, 5,298,633, 5,298,522, 5,298,498,5,290,800, 5,290,788, 5,284,949, 5,280,045, 5,270,319, 5,266,562,5,256,680, 5,250,700, 5,250,552, 5,248,682, 5,244,917, 5,240,929,5,234,939, 5,234,937, 5,232,939, 5,225,571, 5,225,418, 5,220,025,5,212,189, 5,212,172, 5,208,250, 5,204,365, 5,202,350, 5,196,431,5,191,084, 5,187,175, 5,185,326, 5,183,906, 5,177,079, 5,171,864,5,169,963, 5,155,122, 5,143,929, 5,143,928, 5,143,927, 5,124,455,5,124,347, 5,114,958, 5,112,846, 5,104,656, 5,098,613, 5,095,037,5,095,019, 5,086,064, 5,081,261, 5,081,147, 5,081,126, 5,075,330,5,066,668, 5,059,602, 5,043,457, 5,037,835, 5,037,811, 5,036,088,5,013,850, 5,013,751, 5,013,736, 500,654, 4,992,448, 4,992,447,4,988,733, 4,988,728, 4,981,865, 4,962,119, 4,959,378, 4,954,519,4,945,099, 4,942,236, 4,931,457, 4,927,835, 4,912,248, 4,910,192,4,904,786, 4,904,685, 4,904,674, 4,904,671, 4,897,397, 4,895,953,4,891,370, 4,870,210, 4,859,686, 4,857,644, 4,853,392, 4,851,412,4,847,303, 4,847,290, 4,845,242, 4,835,166, 4,826,990, 4,803,216,4,801,598, 4,791,129, 4,788,205, 4,778,818, 4,775,679, 4,772,703,4,767,776, 4,764,525, 4,760,051, 4,748,153, 4,725,616, 4,721,712,4,713,393, 4,708,966, 4,695,571, 4,686,235, 4,686,224, 4,680,298,4,678,802, 4,652,564, 4,644,005, 4,632,923, 4,629,793, 4,614,741,4,599,360, 4,596,828, 4,595,694, 4,595,686, 4,594,357, 4,585,755,4,579,866, 4,578,390, 4,569,942, 4,567,201, 4,563,476, 4,559,348,4,558,067, 4,556,672, 4,556,669, 4,539,326, 4,537,903, 4,536,503,4,518,608, 4,514,415, 4,512,990, 4,501,755, 4,495,197, 4,493,839,4,465,687, 4,440,779, 4,440,763, 4,435,420, 4,412,995, 4,400,534,4,355,034, 4,335,141, 4,322,420, 4,275,064, 4,244,963, 4,235,908,4,234,593, 4,226,887, 4,201,778, 4,181,720, 4,173,650, 4,173,634,4,145,444, 4,128,664, 4,125,612, 4,124,726, 4,124,707, 4,117,135,4,027,031, 4,024,284, 4,021,553, 4,021,550, 4,018,923, 4,012,527,4,011,326, 3,998,970, 3,998,954, 3,993,763, 3,991,212, 3,984,405,3,978,227, 3,978,219, 3,978,202, 3,975,543, 3,968,224, 3,959,368,3,949,082, 3,949,081, 3,947,475, 3,936,450, 3,934,018, 3,930,005,3,857,955, 3,856,962, 3,821,377, 3,821,401, 3,789,121, 3,789,123,3,726,978, 3,694,471, 3,691,214, 3,678,169, 3,624,216;

immunosuppressive agents, as disclosed in U.S. Pat. Nos. 4,450,159,4,450,159, 5,905,085, 5,883,119, 5,880,280, 5,877,184, 5,874,594,5,843,452, 5,817,672, 5,817,661, 5,817,660, 5,801,193, 5,776,974,5,763,478, 5,739,169, 5,723,466, 5,719,176, 5,696,156, 5,695,753,5,693,648, 5,693,645, 5,691,346, 5,686,469, 5,686,424, 5,679,705,5,679,640, 5,670,504, 5,665,774, 5,665,772, 5,648,376, 5,639,455,5,633,277, 5,624,930, 5,622,970, 5,605,903, 5,604,229, 5,574,041,5,565,560, 5,550,233, 5,545,734, 5,540,931, 5,532,248, 5,527,820,5,516,797, 5,514,688, 5,512,687, 5,506,233, 5,506,228, 5,494,895,5,484,788, 5,470,857, 5,464,615, 5,432,183, 5,431,896, 5,385,918,5,349,061, 5,344,925, 5,330,993, 5,308,837, 5,290,783, 5,290,772,5,284,877, 5,284,840, 5,273,979, 5,262,533, 5,260,300, 5,252,732,5,250,678, 5,247,076, 5,244,896, 5,238,689, 5,219,884, 5,208,241,5,208,228, 5,202,332, 5,192,773, 5,189,042, 5,169,851, 5,162,334,5,151,413, 5,149,701, 5,147,877, 5,143,918, 5,138,051, 5,093,338,5,091,389, 5,068,323, 5,068,247, 5,064,835, 5,061,728, 5,055,290,4,981,792, 4,810,692, 4,410,696, 4,346,096, 4,342,769, 4,317,825,4,256,766, 4,180,588, 4,000,275, 3,759,921;

analgesic agents, as disclosed in U.S. Pat. Nos. 5,292,736, 5,688,825,5,554,789, 5,455,230, 5,292,736, 5,298,522, 5,216,165, 5,438,064,5,204,365, 5,017,578, 4,906,655, 4,906,655, 4,994,450, 4,749,792,4,980,365, 4,794,110, 4,670,541, 4,737,493, 4,622,326, 4,536,512,4,719,231, 4,533,671, 4,552,866, 4,539,312, 4,569,942, 4,681,879,4,511,724, 4,556,672, 4,721,712, 4,474,806, 4,595,686, 4,440,779,4,434,175, 4,608,374, 4,395,402, 4,400,534, 4,374,139, 4,361,583,4,252,816, 4,251,530, 5,874,459, 5,688,825, 5,554,789, 5,455,230,5,438,064, 5,298,522, 5,216,165, 5,204,365, 5,030,639, 5,017,578,5,008,264, 4,994,450, 4,980,365, 4,906,655, 4,847,290, 4,844,907,4,794,110, 4,791,129, 4,774,256, 4,749,792, 4,737,493, 4,721,712,4,719,231, 4,681,879, 4,670,541, 4,667,039, 4,658,037, 4,634,708,4,623,648, 4,622,326, 4,608,374, 4,595,686, 4,594,188, 4,569,942,4,556,672, 4,552,866, 4,539,312, 4,536,512, 4,533,671, 4,511,724,4,440,779, 4,434,175, 4,400,534, 4,395,402, 4,391,827, 4,374,139,4,361,583, 4,322,420, 4,306,097, 4,252,816, 4,251,530, 4,244,955,4,232,018, 4,209,520, 4,164,514 4,147,872, 4,133,819, 4,124,713,4,117,012, 4,064,272, 4,022,836, 3,966,944;

cholinergic agents, as disclosed in U.S. Pat. Nos. 5,219,872, 5,219,873,5,073,560, 5,073,560, 5,346,911, 5,424,301, 5,073,560, 5,219,872,4,900,748, 4,786,648, 4,798,841, 4,782,071, 4,710,508, 5,482,938,5,464,842, 5,378,723, 5,346,911, 5,318,978, 5,219,873, 5,219,872,5,084,281, 5,073,560, 5,002,955, 4,988,710, 4,900,748, 4,798,841,4,786,648, 4,782,071, 4,745,123, 4,710,508;

adrenergic agents, as disclosed in U.S. Pat. Nos. 5,091,528, 5,091,528,4,835,157, 5,708,015, 5,594,027, 5,580,892, 5,576,332, 5,510,376,5,482,961, 5,334,601, 5,202,347, 5,135,926, 5,116,867, 5,091,528,5,017,618, 4,835,157, 4,829,086, 4,579,867, 4,568,679, 4,469,690,4,395,559, 4,381,309, 4,363,808, 4,343,800, 4,329,289, 4,314,943,4,311,708, 4,304,721, 4,296,117, 4,285,873, 4,281,189, 4,278,608,4,247,710, 4,145,550, 4,145,425, 4,139,535, 4,082,843, 4,011,321,4,001,421, 3,982,010, 3,940,407, 3,852,468, 3,832,470;

antihistamine agents, as disclosed in U.S. Pat. Nos. 5,874,479,5,863,938, 5,856,364, 5,770,612, 5,702,688, 5,674,912, 5,663,208,5,658,957, 5,652,274, 5,648,380, 5,646,190, 5,641,814, 5,633,285,5,614,561, 5,602,183, 4,923,892, 4,782,058, 4,393,210, 4,180,583,3,965,257, 3,946,022, 3,931,197;

steroidal agents, as disclosed in U.S. Pat. Nos. 5,863,538, 5,855,907,5,855,866, 5,780,592, 5,776,427, 5,651,987, 5,346,887, 5,256,408,5,252,319, 5,209,926, 4,996,335, 4,927,807, 4,910,192, 4,710,495,4,049,805, 4,004,005, 3,670,079, 3,608,076, 5,892,028, 5,888,995,5,883,087, 5,880,115, 5,869,475, 5,866,558, 5,861,390, 5,861,388,5,854,235, 5,837,698, 5,834,452, 5,830,886, 5,792,758, 5,792,757,5,763,361, 5,744,462, 5,741,787, 5,741,786, 5,733,899, 5,731,345,5,723,638, 5,721,226, 5,712,264, 5,712,263, 5,710,144, 5,707,984,5,705,494, 5,700,793, 5,698,720, 5,698,545, 5,696,106, 5,677,293,5,674,861, 5,661,141, 5,656,621, 5,646,136, 5,637,691, 5,616,574,5,614,514, 5,604,215, 5,604,213, 5,599,807, 5,585,482, 5,565,588,5,563,259, 5,563,131, 5,561,124, 5,556,845, 5,547,949, 5,536,714,5,527,806, 5,506,354, 5,506,221, 5,494,907, 5,491,136, 5,478,956,5,426,179, 5,422,262, 5,391,776, 5,382,661, 5,380,841, 5,380,840,5,380,839, 5,373,095, 5,371,078, 5,352,809, 5,344,827, 5,344,826,5,338,837, 5,336,686, 5,292,906, 5,292,878, 5,281,587, 5,272,140,5,244,886, 5,236,912, 5,232,915, 5,219,879, 5,218,109, 5,215,972,5,212,166, 5,206,415, 5,194,602, 5,166,201, 5,166,055, 5,126,488,5,116,829, 5,108,996, 5,099,037, 5,096,892, 5,093,502, 5,086,047,5,084,450, 5,082,835, 5,081,114, 5,053,404, 5,041,433, 5,041,432,5,034,548, 5,032,586, 5,026,882, 4,996,335, 4,975,537, 4,970,205,4,954,446, 4,950,428, 4,946,834, 4,937,237, 4,921,846, 4,920,099,4,910,226, 4,900,725, 4,892,867, 4,888,336, 4,885,280, 4,882,322,4,882,319, 4,882,315, 4,874,855, 4,868,167, 4,865,767, 4,861,875,4,861,765, 4,861,763, 4,847,014, 4,774,236, 4,753,932, 4,711,856,4,710,495, 4,701,450, 4,701,449, 4,689,410, 4,680,290, 4,670,551,4,664,850, 4,659,516, 4,647,410, 4,634,695, 4,634,693, 4,588,530,4,567,000, 4,560,557, 4,558,041, 4,552,871, 4,552,868, 4,541,956,4,519,946, 4,515,787, 4,512,986, 4,502,989, 4,495,102; the disclosuresof which are herein incorporated by reference.

Also of interest are analogs of the above compounds. For all of theabove active agents, the active agents may be present aspharmaceutically acceptable salts, as mentioned above.

Delivery Vehicle Component

The delivery compositions of the invention are compositions that areformulated for delivery of an active agent to a topical location, suchas a keratinized skin surface or a mucosal surface of a mammaliansubject, such as a human subject. By keratinized skin surface is meant askin location of a subject, i.e., a location of the external covering orintegument of an animal body. By mucosal surface is meant a location ofa subject that includes a mucosal membrane, such as the inside of themouth, in the inside of the nose, etc.

Because the dermal delivery formulations of the invention are formulatedfor delivery to topical location, they are formulated so as to bephysiologically compatible with the topical location for which they areformulated. Accordingly, when contacted with the target keratinized skinsurface or mucosal surface for which they are formulated, the deliverycompositions do not cause substantial, if any, physiological responses(such as inflammation or irritation) that would render the use of thedelivery compositions unsuitable for topical application.

The delivery compositions of the invention include an amount of theactive agent-calcium phosphate particle complexes included in a deliveryvehicle component. The delivery vehicle component refers to that portionof the delivery composition that is not the active agent-calciumphosphate particle complex component.

The delivery vehicle component of the delivery compositions of theinvention may vary, as desired, where the particular ingredients of agiven delivery vehicle component will depend, at least in part, on thenature of the particular composition. Delivery compositions of interestinclude liquid formulations, such as lotions (liquids containinginsoluble material in the form of a suspension or emulsion, intended forexternal application, including spray lotions) and aqueous solutions,semi-solid formulations, such as gels (colloids in which the dispersephase has combined with the dispersion medium to produce a semisolidmaterial, such as a jelly), creams (soft solids or thick liquids) andointments (soft, unctuous preparations), and solid formulations, such astopical patches. As such, delivery vehicle components of interestinclude, but are not limited to: emulsions of the oil-in-water (O/W) andthe water in-oil (W/O) type, milk preparations, lotions, creams,ointments, gels, serum, powders, masks, packs, sprays, aerosols orsticks

Lotions

Lotions are liquid compositions where the viscosity is 50,000 cP orless, such as 10,000 cP or less, as determined using a RotationalViscometer, which Measures viscosity by measuring the running torque ofthe cylindrical rotors immersed in a sample, viscosity determinationprotocol at a temperature of 25° C., as described in JIS K 7117: TestingMethods For Viscosity With A Rotational Viscometer Of Resins In TheLiquid or ASTM D 2196-86: Test Methods for Rheological Properties onNon-Newtonian Materials by Rotational (Brookfield) Viscometer.

Lotion delivery vehicle components of interest may include a number ofdifferent ingredients, including but not limited to: water, emollients,natural oils, silicone oils, thickening agents or viscosity modifiers,synthetic or natural esters, fatty acids, alcohols, humectants,emulsifiers, preservative systems, colorants, fragrances, etc. Amountsof these materials may range from 0.001 to 99%, such as from 0.1 to 50%,including from 1 to 20% by weight of the composition, as desired.

Emollients are compounds that replace or add to lipids and natural oilsin the skin. The term emollient, as used herein, is intended to includeconventional lipid materials (e.g., fats, waxes, and other waterinsoluble materials), polar lipids (e.g., lipid materials which havebeen modified to render them more water soluble), silicones andhydrocarbons. Emollients of interest include, but are not limited to:diisopropyl adipate, isopropyl myristate, isopropyl palmitate,ethylhexyl palmitate, isodecyl neopentanoate, C₁₂₋₁₅ alcohols benzoate,diethylhexyl maleate, PPG-14 butyl ether, PPG-2 myristyl etherpropionate, cetyl ricinoleate, cholesterol stearate, cholesterolisosterate, cholesterol acetate, jojoba oil, cocoa butter, shea butter,lanolin, and lanolin esters.

Silicone oils may be divided into the volatile and non-volatile variety.The term “volatile” as used herein refers to those materials which havea measurable vapor pressure at ambient temperature. Volatile siliconeoils of interest include but are not limited to: cyclic or linearpolydimethylsiloxanes containing from 3 to 9, such as from 4 to 5,silicon atoms. Linear volatile silicone materials may have viscositiesof 5 centistokes or less at 25° C., while cyclic materials may haveviscosities of 10 centistokes or less. Nonvolatile silicone oils ofinterest include, but are not limited to: polyalkyl siloxanes,polyalkylaryl siloxanes and polyether siloxane copolymers. Theessentially non-volatile polyalkyl siloxanes of interest include, forexample, polydimethyl siloxanes with viscosities of 5 to 100,000centistokes at 25° C.

Suitable esters include, but are not limited to: alkenyl or alkyl estersof fatty acids having 10 to 20 carbon atoms, such as isopropylpalmitate, isopropyl isostearate, isononyl isonanonoate, oleylmyristate, oleyl stearate, and oleyl oleate; ether-esters, such as fattyacid esters of ethoxylated fatty alcohols; polyhydric alcohol esters;ethylene glycol mono and di-fatty acid esters; diethylene glycol mono-and di-fatty acid esters, polyethylene glycol (200-6000) mono- anddi-fatty acid esters; propylene glycol mono- and di-fatty acid esters,such as polypropylene glycol 2000 monooleate, polypropylene glycol 2000monostearate, ethoxylated propylene glycol monostearate; glyceryl mono-and di-fatty acid esters; polyglycerol poly-fatty esters, such asethoxylated glyceryl monostearate, 1,3-butylene glycol monostearate,1,3-butylene glycol distearate; polyoxyethylene polyol fatty acid ester;sorbitan fatty acid esters; and polyoxyethylene sorbitan fatty acidesters are satisfactory polyhydric alcohol esters; wax esters such asbeeswax, spermaceti, myristyl myristate, stearyl stearate; sterolsesters, of which soya sterol and cholesterol fatty acid esters areexamples thereof. Both vegetable and animal sources of these compoundsmay be used. Examples of such oils include, but are not limited to:castor oil, lanolin oil, C10-18 triglycerides, caprylic/caprictriglycerides, sweet almond oil, apricot kernel oil, sesame oil,camelina sativa oil, tamanu seed oil, coconut oil, corn oil, cottonseedoil, linseed oil, ink oil, olive oil, palm oil, illipe butter, rapeseedoil, soybean oil, grapeseed oil, sunflower seed oil, walnut oil, and thelike. Also suitable are synthetic or semi-synthetic glyceryl esters,such as fatty acid mono-, di-, and triglycerides which are natural fatsor oils that have been modified, for example, mono-, di- or triesters ofpolyols such as glycerin. In an example, a fatty (C₁₂₋₂₂) carboxylicacid is reacted with one or more repeating glyceryl groups. glycerylstearate, diglyceryl diiosostearate, polyglyceryl-3 isostearate,polyglyceryl-4 isostearate, polyglyceryl-6 ricinoleate, glyceryldioleate, glyceryl diisotearate, glyceryl tetraisostearate, glyceryltrioctanoate, diglyceryl distearate, glyceryl linoleate, glycerylmyristate, glyceryl isostearate, PEG castor oils, PEG glyceryl oleates,PEG glyceryl stearates, and PEG glyceryl tallowates.

Fatty acids of interest include, but are not limited to: those havingfrom 10 to 30 carbon atoms, such as pelargonic, lauric, myristic,palmitic, stearic, isostearic, hydroxystearic, oleic, linoleic,ricinoleic, arachidic, behenic and erucic acids.

Humectants of the polyhydric alcohol-type may also find use in thecompositions, where examples of polyhydric alcohols include, but are notlimited to: glycerol (also known as glycerin), polyalkylene glycols,alkylene polyols and their derivatives, including propylene glycol,dipropylene glycol, polypropylene glycol, polyethylene glycol andderivatives thereof, sorbitol, hydroxypropyl sorbitol, hexylene glycol,1,3-butylene glycol, 1,2,6-hexanetriol, ethoxylated glycerol,propoxylated glycerol and mixtures thereof. Also of interest are sugars,e.g., glucose, fructose, honey, hydrogenated honey, inositol, maltose,mannitol, maltitol, sorbitol, sucrose, xylitol, xylose, etc. Whenpresent, the amount of humectant may range from 0.001 to 25%, such asfrom about 0.005 to 20%, including from about 0.1 to 15%, where in someinstances the amount of humectant ranges from 0.5 to 30%, such asbetween 1 and 15% by weight of the composition.

Emulsifiers may also be present in the vehicle compositions. Whenpresent, the total concentration of the emulsifier may range from 0.01to 40%, such as from 1 to 20%, including from 1 to 5% by weight of thetotal composition. Emulsifiers of interest include, but are not limitedto: anionic, nonionic, cationic and amphoteric actives.

Nonionic surfactants of interest include those with a C₁₀-C₂₀ fattyalcohol or acid hydrophobe condensed with from about 2 to about 100moles of ethylene oxide or propylene oxide per mole of hydrophobe;C₂-C₁₀ alkyl phenols condensed with from 2 to 20 moles of alkyleneoxide; mono- and di-fatty acid esters of ethylene glycol; fatty acidmonoglyceride; sorbitan, mono- and di-C₈-C₂O fatty acids; andpolyoxyethylene sorbitan as well as combinations thereof. Alkylpolyglycosides and saccharide fatty amides (e.g. methyl gluconamides)are also of interest nonionic emulsifiers. Anionic emulsifiers ofinterest include soap, alkyl ether sulfate and sulfonates, alkylsulfates and sulfonates, alkylbenzene sulfonates, alkyl and dialkylsulfosuccinates, C₈-C₂₀ acyl isethionates, C₈-C₂₀ alkyl etherphosphates, alkylethercarboxylates and combinations thereof.

Where desired, preservatives can include in the compositions, e.g., toprotect against the growth of potentially harmful microorganisms.Preservatives of interest include alkyl esters of para-hydroxybenzoicacid, hydantoin derivatives, propionate salts, and a variety ofquaternary ammonium compounds. Specific preservatives of interestinclude, but are not limited to: iodopropynyl butyl carbamate,phenoxyethanol, methyl paraben, propyl paraben, imidazolidinyl urea,sodium dehydroacetate, benzyl alcohol, benzylhemiformal, benzylparaben,5-bromo-5-nitro-1,3-dioxane, 2-bromo-2-nitropropane-1,3-diol, caprylylglycol, ethylhexylglycerin, phenoxyethanol sorbic acid, methylparaben,propylparaben, ethylpareben, butylparaben, sodium benzoate, potassiumsorbate, disodium salt of ethylenediaminetetraacetic acid,chloroxylenol, DMDM Hydantoin, 3-iodo-2-propylbutyl carbamate,chlorhexidine digluconate, phenoxyethanol, diazolidinyl urea, biguanidederivatives, calcium benzoate, calcium propionate, caprylyl glycol,biguanide derivatives, captan, chlorhexidine diacetate, chlorhexidinedigluconate, chlorhexidine dihydrochloride, chloroacetamide,chlorobutanol, p-chloro-m-cresol, chlorophene, chlorothymol,chloroxylenol, m-cresol, o-cresol, DEDM Hydantoin, DEDM Hydantoindilaurate, dehydroacetic acid, diazolidinyl urea, dibromopropamidinediisethionate, DMDM Hydantoin, and the like When present, preservativesmay be present in the delivery compositions in amounts ranging fromabout 0.01% to about 10% by weight of the composition.

Thickening agents or viscosity modifiers may be included in the deliverycompositions. Thickening agents of interest include, but are not limitedto: polysaccharides, such as starches, natural/synthetic gums andcellulosics. Starches of interest include, but are not limited to,chemically modified starches, such as aluminum starch octenylsuccinate.Gums of interest include, but are not limited to: xanthan, sclerotium,pectin, karaya, arabic, agar, guar, carrageenan, alginate andcombinations thereof. Suitable cellulosics include, but are not limitedto: hydroxypropyl cellulose, hydroxypropyl methylcellulose,ethylcellulose and sodium carboxy methylcellulose. Synthetic polymersare still a further class of effective thickening agent. This categoryincludes crosslinked polyacrylates such as the Carbomers andpolyacrylamides such as Sepigel® 305. When present, amounts of thethickener may range from 0.001 to 5%, such as from 0.1 to 2%, includingfrom 0.2 to 0.5% by weight.

In some instances, natural or synthetic organic waxes may be present,e.g., one or more natural or synthetic waxes such as animal, vegetable,or mineral waxes. In some instances, such waxes will have a meltingpoint ranging from 20 to 150° C., such as from 30 to 100° C., including35 to 75° C. Examples of such waxes include waxes such as polyethyleneor synthetic wax; or various vegetable waxes such as bayberry,candelilla, ozokerite, acacia, beeswax, ceresin, cetyl esters, flowerwax, citrus wax, carnauba wax, jojoba wax, japan wax, polyethylene,microcrystalline, rice bran, lanolin wax, mink, montan, bayberry,ouricury, ozokerite, palm kernel wax, paraffin, avocado wax, apple wax,shellac wax, clary wax, spent grain wax, grape wax, and polyalkyleneglycol derivatives thereof such as PEG6-20 beeswax, or PEG-12 carnaubawax; or fatty acids or fatty alcohols, including esters thereof, such ashydroxystearic acids (for example 12-hydroxy stearic acid), tristearin,and tribehenin. Also of interest are Acrocomia Aculeata Seed Butter,Almond Butter, Aloe Butter, Apricot Kernel Butter, Argan Butter, AttaleaMaripa Seed Butter, Avocado Butter, Babassu Butter, Bacuri Butter,Bagura Soft Butter, Baobab Soft Butter, Bassia Butyracea Seed Butter,Bassia Latifolia Seed Butter, Black Currant Seed Butter, Brazil NutButter, Camelina Butter, Camellia Butter, Candelilla Butter, CarnaubaButter, Carpotroche Brasiliensis Seed Butter, Chamomile Butter, CocoaButter, Coconut Butter, Coffee Butter, Cotton Soft Butter, CranberryButter, Cupuacu Butter, Grape Seed Butter, Hazel Nut Butter, Hemp SeedButter, Horsetail Butter, Illipe Butter, Irvingia Gabonensis KernelButter, Jojoba Butter, Karite Butter, Kokum Butter, Kukui Butter,Lavender Butter, Lemon Butter, Lime Butter, Macadamia Butter, MangoButter, Marula Butter, Monoi Butter, Mowrah Butter, Mucaja Butter,Murumuru Butter, Olea Butter, Olive Butter, Orange Butter, Palm Oil,Passion Butter, Phulwara Butter, Pistachio Butter, Pomegranate Butter,Pumpkin Butter, Raspberry Butter, Rice Butter, Sal Butter, SapucainhaButter, Seasame Butter, Shea Butter, Soy Butter Tamanu Butter, SunflowerSeed Butter, Sweet almond Butter, Tangerine Butter, Tucuma Seed Butter,Ucuuba Butter and Wheat Germ Butter.

Colorants, fragrances and abrasives may also be included in the deliverycompositions. Each of these substances may range from 0.05 to 5%, suchas from 0.1 and 3% by weight. Colorants of interest include titaniumdioxide, where appropriate surface-treated (codified in the Color Indexunder the reference CI 77,891), manganese violet (CI 77,742),ultramarine blue (CI 77,007), chromium oxide (CI 77,288), hydratedchromium oxide (CI 77,289), ferric blue (CI 77,510), zinc oxide,zirconium dioxide. Specific colorants of interest include: D & C red no.19 (CI 45,170), D & C red no. 9 (CI 15,585), D & C red no. 21 (CI45,380), D & C orange no. 4 (CI 15,510), D & C orange no. 5 (CI 45,370),D & C red no. 27 (CI 45,410), D & C red no. 13 (CI 15,630), D & C redno. 7 (CI 15,850:1), D & C red no. 6 (CI 15,850:2), D & C yellow no. 5(CI 19,140), D & C red no. 36 (CI 12,085), D & C orange no. 10 (CI45,425), D & C yellow no. 6 (CI 15,985), D & C red no. 30 (CI 73,360), D& C red no. 3 (CI 45,430), carbon black (CI 77,266), cochineal carminelake (CI 75,470), natural or synthetic melanin, and aluminium lakes.

Fragrances of interest include: Abies Alba Leaf Oil, Acetaldehyde,Acetanilid, Acetic Acid, Achillea Millefolium Oil, Actinidia Chinensis(Kiwi) Fruit Water, Adipic Acid, Agar, Alcohol Denat., Algin, AloeBarbadensis Leaf, Amyl Acetate, Amyl Benzoate, Amyl Cinnamal, Anethole,Anise alcohol, Anthemis Nobilis Flower Water, Benzaldehyde, BenzylAlcohol, Betula Alba Oil, Boswellia Serrata Oil, Butyl Acetate, ButylLactate, Calendula Officinalis Flower Oil, Camellia Sinensis Leaf Water,Camphor, Capsaicin, Cedrol, Cinnamal, Citral, Citronellol, CitrusAurantifolia (Lime) Oil, Citrus Aurantium Dulcis (Orange) Oil, CitrusGrandis (Grapefruit) Oil, Citrus Tangerina (Tangerine) Peel Oil,Coumarin, Diacetone Alcohol, Ethyl Cinnamate, Ethyl Ether, EucalyptusCaryophyllus (Clove) Flower Oil, Farnesol, Gardenia Florida Oil,Geranium Maculatum Oil, Hexyl Cinnamal, Hydrogenated Rosin, IlliciumVerum (Anise) Oil, Isoamyl Acetate, Juniperus Mexicana Oil, LaurusNobilis Oil, Lavandula angustifolia (Lavender) Oil, MelaleucaAlternifolia (Tea Tree) Leaf Oil, Melissa Officinalis Leaf Oil, MenthaPiperita (Peppermint) Oil, Menthol, 2-Naphthol, Origanum Majorana LeafOil, Panax Ginseng Root Extract, Pelargonic Acid, Pelargonim GraveolensFlower Oil, Pinus Silvestris Cone Oil, Prunus Armeniaca (Apricot) KernelOil, Rosa Canina Flower Oil, Rosmarinus Officinalis (Rosemary) Leaf Oil,Santalum Album (Sandalwood) Oil, Thymus Vugaris (Thyme) Oil, Vanillin,Vitis Vinifera (Grape) Leaf Oil, Zingiber Officinale (Ginger) Root Oil.

Semi-Solid Delivery Compositions

Also of interest are semi-solid delivery compositions, such as gels,creams and ointments. Such compositions may be mixtures of (in additionto the active agent-calcium phosphate particle complex) water, watersoluble polymers, preservatives, alcohols, polyvalent alcohols,emulsifying agents, humectants, wax, solvents, thickeners, plasticizers,pH regulators, water-retaining agents and the like. Furthermore, suchcompositions may also contain other physiologically acceptableexcipients or other minor additives, such as fragrances, dyes,emulsifiers, buffers, antibiotics, stabilizers or the like. Examples ofthese types of compounds are provided above.

Topical Patches

Also of interest are solid formulations, such as topical patchformulations. Topical patch formulations may vary significantly. Topicalpatch formulations may include an active agent layer, a support and arelease liner. The active agent layer may include an amount of theactive agent-particle complexes in a matrix, where the matrix mayinclude one or more of: adhesives, such as pressure sensitive rubber andacrylic acids, hydrogels, physiologically acceptable excipients or otherminor additives, such as fragrances, dyes, emulsifiers, buffers,antibiotics, stabilizers or the like. The support may be made of aflexible material which is capable of fitting in the movement of humanbody and includes, for example, plastic films, various non-wovenfabrics, woven fabrics, spandex, and the like. Various inert coveringsmay be employed, which include the various materials which may find usein plasters, described below. Alternatively, non-woven or wovencoverings may be employed, particularly elastomeric coverings, whichallow for heat and vapor transport. These coverings allow for cooling ofthe pain site, which provides for greater comfort, while protecting thegel from mechanical removal. The release liner may be made of anyconvenient material, where representative release films includepolyesters, such as PET or PP, and the like.

Aerosol Compositions

Also of interest are aerosol compositions formulations to beadministered via inhalation. These aerosol formulations can be placedinto pressurized acceptable propellants, such asdichlorodifluoromethane, propane, nitrogen, and the like. They may alsobe formulated for non-pressured preparations, such as for use in anebulizer or an atomizer. In some embodiments, the formulations arepowdered aerosol formulations which include the active agent boundparticles suspended or dispersed in a propellant or a propellant andsolvent. The propellant may be a mixture of liquefiedchlorofluorocarbons (CFCs) which are selected to provide the desiredvapor pressure and stability of the formulation. Propellants 11, 12 and114 are the most widely used propellants in aerosol formulations forinhalation administration. Other commonly used propellants includePropellants 113, 142b, 152a, 124, and dimethyl ether. The compound1,1,1,2-tetrafluoroethane is also a commonly used propellant formedicinal aerosol formulations. The propellant may be 40 to 90% byweight of the total inhalation composition.

Methods of Making Delivery Compositions

Aspects of the invention further include methods of making the deliverycompositions. While any convenient fabrication protocol may be employed,in some instances fabrication protocols include first preparing theactive agent-particle complexes. Following production of the activeagent-particle complexes, the resultant complexes are then combinedwithin the delivery composition component using any convenient protocol.

The active agent-particle complexes may be produced using any convenientprotocol. One protocol of interest includes first producing a liquidcomposition of the active agent, such as an aqueous composition of theactive agent, and then combining the liquid composition with an amountof uniform, rigid, spherical, nanoporous calcium phosphate particles(with agitation where desired) under conditions sufficient to producethe desired active agent-particle complexes. As such, in certainembodiments a fluid composition of unbound particles, e.g., a slurry ofunbound particles in a suitable solvent system (such as an aqueous ornon-aqueous solvent system) is combined with a suitable amount of activeagent.

Particle Pre-treatment

As desired, the unbound particles may be pretreated in some manner priorto combination with the active agent. As such, preparation of the activeagent bound particles may include a pre-treatment step, such as aninitial pH adjustment step. In this step, the unbound particles aremodified by contacting them with one or more agents, such as a pHadjustment agent, in order to provide for desired active agent bindingto the particles. The particular nature of the pH adjustment, ifemployed, varies depending on the type of active agent that is to bebound to the particles. One category of active agents of interest arethose that include acidic and/or basic charged moieties and a molecularmass greater than a few thousand Daltons, e.g., having a mass of 3000Daltons or greater, such as 5,000 Daltons or greater, e.g., 10,000Daltons or greater, 25,000 Daltons or greater, 50,000 Daltons orgreater, 75,000 Daltons or greater, 100,000 Daltons or greater, 250,000Daltons or greater, 500,000 Daltons or greater, 750,000 Daltons orgreater, 1,000,000 Daltons or greater. Examples of such active agentsinclude, but are not limited to proteins, nucleic acids andpolysaccharides. Such active agents may strongly bind to the calciumand/or phosphate sites of the particles under a broad range of pHconditions. Accordingly, for these types of active agents, pHmodification may or may not be performed, as desired. Where pHmodification is desired, pH modification may be performed by using anyconvenient pH adjustment agent, e.g., an acid or alkaline agent. pHadjustment agents of interest include, but are not limited to: lacticacid, glycolic acid, triethanolamine and sodium hydroxide. In someinstances, pH adjustment agents are selected that do not block thecalcium and/or phosphate binding sites of particles.

Another category of active agents of interest are those that includeacidic and/or basic charged moieties and a molecular mass that does notexceed a few thousand Daltons, e.g., having a mass of 2500 Daltons orless, such as 1500 Daltons or less. Examples of such active agentsinclude, but are not limited to organic acid and amine compounds. Suchactive agents bind to the particles at a specific pH. Pretreatment ofthe particles by optimizing the pH, and/or addition of specific ioniccompounds into the binding solution (described in greater detail below)may be employed, as desired.

Yet another category of active agents of interest are water-solublesmall molecules with non-charged or weakly charged moieties. Examples ofsuch compounds include, but are not limited to: saccharides, glycosidesand amino acid derivatives. For this category of active agents, anaqueous and/or organic solvent mixture, such as ethanol/water oracetonitrile/water may be employed for a pretreatment and active agentbinding, as desired.

Yet another category of active agents of interest are water solublesmall molecules with hydrophobic moieties. For such active agents,pretreatment may include contacting the particles with surface modifyingagents, e.g., agents that include one or more charged groups and one ormore hydrophobic tails, such as but not limited to sodium dodecylsulfate, sodium lauryl sulfate and sodium lauryl phosphate, and thelike.

Yet another category of active agents of interest are water insolublemolecules. Examples of water insoluble molecules of interest include,but are not limited to: amino acid derivatives, polyphenols, andretinoids. For such active agents, the use of organic solvents such asethanol and dimethyl sulfoxide (DMSO) as a pretreatment agent and/orloading solvent may be employed, as desired.

In some instances, particles are pretreated with an ionic modificationagent. Ionic modification agents include, but are not limited to,calcium ion modification agents, such as CaCl₂, phosphate ionmodification agents, such as sodium phosphate, etc.

Following any particle pretreatment step, e.g., as described above, insome embodiments the particles are subjected to a washing step. Forexample, in some instances, it may be desirable to remove excess salt orions from the particles by washing, filtering or decanting the particlesprior to active agent binding. Any convenient wash protocol and fluidmay be employed for this step.

Complex Formation

Following any pretreatment and/or washing, such as described above (ifnecessary), the unbound particles are combined with active agent toproduce active agent bound particles. The active agent can be either inpowder or solution form, as desired. Any suitable protocol for combiningthe active agent and the particles may be employed, such as simplestatic mixing in a vessel, etc. The pH of the composition during bindingmay be selected to provide for maximum binding, e.g., by employing a pHadjustment agent, such as described above. For example, in someinstances basic active agents are combined with the particles underbasic conditions and acidic active agents are combined with theparticles under acidic conditions. Therefore, the pH of the complexformation reaction may range, in some instances from 5 to 14. In certaininstances, the pH is 10 or less, where, depending on the length of timeemployed for complex formation, the pH may be selected so as to avoidsubstantial particle degradation, e.g., may be selected to be 5.2 orgreater.

As indicated above, any convenient solvent system may be employed forproducing the active agent-particle complexes. As indicated above, thesolvent system employed in binding the active agent to the particles toproduce the active agent-particle complexes may vary. Solvent systemsfinding use in preparing the active agent-particle complexes may be madeup of a single solvent or a plurality of two or more different solvents.Solvents that are present in solvent systems of interest may be polar(i.e., they have a dielectric constant of 15 or greater) or non-polar(i.e., they have a dielectric constant of less than 15), and protic(such that they solvate anions (negatively charged solutes) strongly viahydrogen bonding) or aprotic (i.e., they have sufficiently large dipolemoments to solvate positively charged species via their dipole).

Protic solvents of interest include, but are not limited to: alcohols,such as methanol, ethanol, propanol, isopropyl alcohol, butanol,pentanol, hexanol, heptanol, octanol, trifluoroethanol, phenol, benzylalcohol, glycerin, ethylene glycol, diethylene glycol; carboxylicacids/amides, such as formic acid, acetic acid, lactic acid, propionicacid, trifluoroacetic acid, formamide; amines, such as ammonia,diethylamine, butyl amine, propyl amine; and water. Aprotic solvents ofinterest include, but are not limited to: hydrocarbons, such as pentane,hexanes, heptane, cyclohexane, methyl cyclohexane, decalin;ketones/aldehydes, such as acetone, methyl ethyl ketone (MEK), methylisobutyl ketone, butanone, pentanone, cyclohexanone, benzaldehyde;aromatic compounds, such as benzene, toluene, trifluorotoluene, xylene,anisole, chlorobenzene, aniline, N,N-dimethylaniline, benzonitrile;ethers, such as dimethoxyethane, dimethyl ether, diethyl ether,diisopropyl ether, methyl t-butyl ether (MTBE), tetrahydrofuran,dioxane, glyme, diglyme, polyethylene glycol (PEG), PEG esters, PEGsorbitans, PEG ethers, PEG esters, polypropylene glycol (PPG), PPGesters, alkoxylated linear alkyl diols, alkoxylated alkyl glucose ether,PPG alkyl ethers; esters/amides, such as methyl acetate, ethyl acetate,propyl acetate, butyl acetate, amyl acetate, ethyl benzoate, benzylbenzoate, dimethyl phthalate, dibutyl phthalate, dimethylacetamide,dimethylformamide (DMF); nitriles, such as acetonitrile; carbonates,such as dimethylcarbonate, diethylcarbonate, propylene carbonate,ethylene carbonate; halogenated compounds, such as carbon tetrachloride,chloroform, dichlormethane, dichloroethane, trichloroethane, freon-11,BMIM-PF₆ ionic liquid; sulfur/Phosphorus-containing compounds, such asdimethyl sulfoxide (DMSO), carbon disulfide, sulfolane,examethylphosphramide; and amines, such as pyridine, triethylamine,N-methylpyrrolidinone (NMP)

Solvents of interest include, but are not limited to: alcohol, alcoholdenat., benzyl glycol, benzyl laurate, benzyllaurate/myristate/palmitate, 1,4-butanediol, 2,3-butanediol, buteth-3,butoxydiglycol, butoxyethanol, butoxyethyl acetate, n-butyl alcohol,t-butyl alcohol, butyl myristate, butylene glycol, butylene glycolpropionate, butyl ethylpropanediyl ethylhexanoate, butyl lactate,butyloctanol, butyloctyl benzoate, butyloctyl salicylate, butylstearate, butylphthalimide, butyrolactone, C₁₂₋₁₅ alkyl benzoate, capricacid, caprylic alcohol, cetearyl octanoate, cetyl stearyl octanoate,chlorobutanol, C₈₋₁₂ acid triglyceride, C₁₂₋₁₈ acid triglyceride, C₉₋₁₂alkane, C₁₀₋₁₃ alkane, C₁₃₋₁₄ alkane, C₁₃₋₁₅ alkane, C₁₄₋₁₇ alkane,C₁₄₋₁₉ alkane, C₁₅₋₁₉ alkane, C₁₅₋₂₃ alkane, C₁₈₋₂₁ alkane, C₈₋₉alkane/cycloalkane, C₉₋₁₀ alkane/cycloalkane, C₉₋₁₁ alkane/cycloalkane,C₉₋₁₆ alkane/cycloalkane, C₁₀₋₁₂ alkane/cycloalkane, C₁₁₋₁₄alkane/cycloalkane, C₁₁₋₁₅ alkane/cycloalkane, C₁₂₋₁₃alkane/cycloalkane, C₈₋₁₀ alkane/cycloalkane/aromatic Hydrocarbons,C₁₂₋₁₅ alkane/cycloalkane/aromatic hydrocarbons, C₉₋₁₀ aromatichydrocarbons, C₁₀₋₁₁ aromatic hydrocarbons, CD alcohol 19, chlorinatedparaffin, C₇₋₈ isoparaffin, C₈₋₉ isoparaffin, C₉₋₁₁ isoparaffin, C₉₋₁₃isoparaffin, C₉₋₁₄ isoparaffin, C₉₋₁₆ isoparaffin, C₁₀₋₁₁ isoparaffin,C₁₀₋₁₂ isoparaffin, C₁₀₋₁₃ isoparaffin, C₂₂₋₁₂ isoparaffin, C₁₁₋₁₃isoparaffin, C₁₁₋₁₄ isoparaffin, C₁₂₋₁₄ isoparaffin, C₁₂₋₂₀ isoparaffin,C₁₃₋₁₄ isoparaffin, C₁₃₋₁₆ isoparaffin, C₂₀₋₄₀ isoparaffin, coixlacryma-jobi (Job's Tears) Seed Water, C₆₋₁₂ perfluoroalkylethanol,C₁₀₋₁₈ triglycerides, cycloethoxymethicone, cycloheptasiloxane,cyclohexanedimethanol, cyclohexasiloxane, cyclomethicone,cyclopentasiloxane, cyclotetrasiloxane, cyclotrisiloxane, decane,1,10-decanediol, decene, decyl alcohol, deodorized kerosene, diacetin,diacetone alcohol, dibutyl adipate, dibutyloctyl malate, dibutyloctylsebacate, dibutyl oxalate, dibutyl phthalate, dibutyl sebacate,di-C12-15 alkyl maleate, diethoxydiglycol, diethoxyethyl succinate,diethylene glycol dibenzoate, diethylhexyl adipate diethylhexyl maleate,diethylhexyl 2,6-naphthalate, diethylhexyl phthalate, diethylhexylsebacate, diethylhexyl succinate, diethyl oxalate, diethyl phthalate,diethyl sebacate, diethyl succinate, diheptylundecyl adipate, dihexyladipate, dihexyldecyl sebacate, diisoamyl malate, diisobutyl adipate,diisobutyl oxalate, diisocetyl adipate, diisodecyl adipate, diisononyladipate, diisooetyl adipate, diisopropyl adipate, diisopropyl oxalate,diisopropyl sebacate, dimethoxydiglycol, dimethyl adipate, dimethylcapramide, dimethyl glutarate, dimethyl isosorbide, dimethyl maleate,dimethyl oxalate, dimethyl phthalate, dimethyl succinate, dimethylsulfone, dioctyl adipate, dioctyl succinate, dioctyldodecyl sebacate,dioxolane, diphenyl methane, di-PPG-3 myristyl ether adipate, dipropyladipate, dipropylene glycol, dipropylene glycol dibenzoate, dipropyleneglycol dimethyl ether, dipropyl oxalate, ditridecyl adipate, dodecene,echium plantagineum seed oil, eicosane, ethoxydiglycol, ethoxydiglycolacetate, ethoxyethanol, ethoxyethanol acetate, ethylene carbonate, ethylether, ethyl hexanediol, ethylhexyl benzoate, ethyl lactate, ethylmacadamiate, ethyl myristate, ethyl oleate, ethyl perfluorobutyl ether,furfural, glycereth-7 benzoate, glycereth-18 benzoate, glycereth-20benzoate, glycereth-7 diisononanoate, glycereth-4,5-lactate, glycereth-5lactate, glycereth-7 lactate, glycereth-7 triacetate, glycine soja(soybean) oil, glycofurol, glycol, hexadecene, hexanediol,1,2-hexanediol, 1,2,6-hexanetriol, hexene, hexyl alcohol, hexyldecylbenzoate, hexyldodecyl salicylate, hexylene glycol, hydrogenatedpolydecene, hydrogenated polydodecene, hydroxymethyl dioxolanone,isoamyl acetate, isobutoxypropanol, isobutyl acetate, isobutyl benzoate,isobutyl stearate, isocetyl salicylate, isodecyl benzoate, isodecylisononanoate, isodecyl octanoate, isodecyl oleate, isododecane,isoeicosane, isohexadecane, isononyl isononanoate, isooctane,isopentane, isopentyldiol, isopropyl acetate, isopropyl citrate,isopropyl laurate, isopropyl myristate, isopropyl palmitate, isopropylphthalimide, isostearyl glycolate, isostearyl stearoyl stearate,laneth-5, lanolin oil, laureth-2 acetate, limonene, 3-methoxybutanol,methoxydiglycol, methoxyethanol, methoxyethanol acetate,methoxyisopropanol, methoxyisopropyl acetate, methoxymethylbutanol,methoxy PEG-7, methoxy PEG-10, methoxy PEG-16, methoxy PEG-25, methoxyPEG-40, methoxy PEG-100, methyl acetate, methylal, methyl benzoate,methylbutenes, methyl gluceth-20 benzoate, methyl hexyl ether, methyllactate, methyl perfluorobutyl ether, methylpropanediol, methylpyrrolidone, methyl soyate, methyl sunflowerseedate, methyltrimethicone, MIBK, mineral oil, mineral spirits, mixed terpenes,momordica grosvenori fruit juice, morpholine, mustelic/palmitictriglyceride, neopentyl glycol, neopentyl glyol dioctanoate,nonocynol-9, octadecane, octadecene, octane, octene, octyl benzoate,octyldodecyl lactate, octyldodecyl octyldodecanoate, octyl isononanoate,octyl isostearate, octyl laurate, octyl palmitate, octyl stearate, oleylalcohol, olive oil PEG-6 esters, peanut oil PEG-6 esters, PBG-33 castoroil, PEG-4, PEG-6, PEG-7, PEG-8, PEG-9, PEG-10, PEG-12, PEG-14, PEG-16,PEG-18, PEG-20, PEG-32, PEG-33, PEG-40, PEG-45, PEG-55, PEG-60, PEG-75,PEG-80, PEG-90, PEG-100, PEG-135, PEG-150, PEG-180, PEG-200, PEG-220,PEG-240, PEG-350, PEG-400, PEG-450, PEG-500, PEG-2 benzyl ether, PEG-15butanediol, PEG-3 methyl ether, PEG-4 methyl ether, PEG-6 methyl ether,PEG-7 methyl ether, PEG-50 glyceryl cocoate, PEG-20 hydrogenated castoroil, PEG/PPG-1/2 copolymer, PEG/PPG-4/2 copolymer, PEG/PPG-5/30copolymer, PEG/PPG-6/2 copolymer, PEG/PPG-7/50 copolymer, PEG/PPG-8/17copolymer, PEG/PPG-10/70 copolymer, PEG/PPG-17/6 copolymer, PEG/PPG-18/4copolymer, PEG/PPG-19/21 copolymer, PEG/PPG-23/17 copolymer,PEG/PPG-23/50 copolymer, PEG/PPG-25/30 copolymer, PEG/PPG-26/31copolymer, PEG/PPG-30/33 copolymer, PEG/PPG-35/9 copolymer, PEG/PPG-38/8copolymer, PEG/PPG-116/66 copolymer, PEG/PPG-125/30 copolymer,PEG/PPG-160/31 copolymer, PEG/PPG-200/70 copolymer, PEG/PPG-240/60copolymer, PEG-10 propylene glycol, 1,5-pentanediol, penetaerythritytetracaprylate/tetracaprate, pentylene glycol, perfluorocaprylylbromide, perfluorodecalin, perfluorodimethylcyclohexane,perfluorohexane, perfluoromethylcyclopentane, perfluoroperhydrobenzyltetralin, perfluoroperhydrophenanthrene, perfluorotetralin, petroleumdistillates, phenoxyisopropanol, phenylpropanol, polyglyceryl-3diisostearate, polyglyceryl-2 dioleate, polyoxyethylene glycoldibenzoate, polyperfluoroethoxymethoxy difluoromethyl ether, PPG-3,PPG-7, PPG-10 butanediol, PPG-2-buteth-3, PPG-3-buteth-5,PPG-5-buteth-7, PPG-7-buteth-4, PPG-7-buteth-10, PPG-12-buteth-16,PPG-15-buteth-20, PPG-20-buteth-30, PPG-20 lanolin alcohol ether, PPG-2myristyl ether propionate, PPG-2 butyl ether, PPG-3 butyl ether,PPG-24-glycereth-24, PPG-25-glycereth-22, PPG-10 glyceryl ether, PPG-55glyceryl ether, PPG-67 glyceryl ether, PPG-70 glyceryl ether, PPG-2methyl ether, PPG-3 methyl ether, PPG-2 methyl ether acetate, PPG-2propyl ether, propanediol, propyl acetate, propylene carbonate,propylene glycol, propylene glycol butyl ether, propylene glycolcaprylate, propylene glycol dibenzoate, propylene glycol methyl ether,propylene glycol myristate, propylene glycol propyl ether, ricinuscommunis (castor) seed oil, SD Alcohol 1, SD Alcohol 3-A, SD Alcohol3-B, SD Alcohol 3-C, SD Alcohol 23-A, SD Alcohol 23-F, SD Alcohol 23-H,SD Alcohol 27-B, SD Alcohol 30, SD Alcohol 31-A, SD Alcohol 36, SDAlcohol 37, SD Alcohol 38-B, SD Alcohol 38-C, SD Alcohol 38-D, SDAlcohol 38-F, SD Alcohol 39, SD Alcohol 39-A, SD Alcohol 39-B, SDAlcohol 39-C, SD Alcohol 39-D, SD Alcohol 40, SD Alcohol 40-A, SDAlcohol 40-B, SD Alcohol 40-C, SD Alcohol 46, sea water, sesamum indicum(sesame) oil, shark liver oil, sorbeth-6, sorbeth-20, sorbeth-30,sorbeth-40, sorbitan trioleate, stearyl benzoate, stearyl heptaroate,tetradecene, tetradecylpropionates, tetrahydrofurfuryl acetate,tetrahydrofurfuryl alcohol, thiolanediol, triacetin, tributyl citrate,tributylcresylbutane, trichloroethane, triethyl phosphate,trimethylhexanol, 2,2,4-timethylpentane, trimethyl pentanol hydroxyethylether.

Where desired, the solvent system can be modified with one or moremodification agents, such as buffers, pH adjusters (acid or base),hydrophilic molecules, hydrophobic molecules or the molecules which haveboth hydrophobic groups and hydrophilic groups (e.g., surfactants).Buffers of interest include, but are not limited to: HCl/sodium citrate,citric acid/sodium citrate, acetic acid/sodium acetate, K₂HPO₄/KH₂PO₄,Na₂HPO₄/NaH₂PO₄, Borax/Sodium hydroxide, as well as biological buffers,e.g., TAPS (3{[tris(hydroxymethyl)methyl]amino}propanesulfonic acid),Bicine (N,N-bis(2-hydroxyethyl)glycine), Tris(tris(hydroxymethyl)methylamine), Tricine(N-tris(hydroxymethyl)methylglycine), HEPES(4-2-hydroxyethyl-1-piperazineethanesulfonic acid), TES(2-[tris(hydroxymethyl)methyl]amino}ethanesulfonic acid), MOPS(3-(N-morpholino)propanesulfonic acid), PIPES(piperazine-N,N-bis(2-ethanesulfonic acid), Cacodylate (dimethylarsinicacid), SSC (saline sodium citrate) and MES(2-(N-morpholino)ethanesulfonic acid); etc.

The components and properties of a particular solvent system, such aspH, composition, temperature, etc., can be selected in view of one ormore properties of the active agent to be complexed with the particles,where such properties may include active agent solubility, structure,pKa, logP, etc.

Following production of the active agent-particle complexes, theresultant complexes are then combined with the delivery compositioncomponent using any convenient protocol. The particular protocolemployed may vary depending on the nature of the delivery compositioncomponent, where in certain instances the delivery composition componentand active agent loaded particles may be combined with mixing to producethe desired delivery composition. While the temperature duringcombination may vary, in some instances the temperature is 80° C., suchas 40° C. or less, such as 30° C. or less, e.g., room temperature orcolder. The amount of active agent-particle complexes that is combinedwith the delivery vehicle may vary. In some embodiments, the amount ofactive agent-particle complexes that is combined with the deliveryvehicle is sufficient to produce a final delivery composition in whichthe amount of active agent-particle complexes ranges from 0.001 to 1000mg/g, such as 0.1 to 200 mg/g and including 1 to 50 mg/g activeagent-particle complexes per gram of delivery composition component. Incertain embodiments, the presence of chelating agents is avoided. The pHof formulation is 5.0 or greater, such as 5.5 or greater.

Utility

The delivery compositions of the invention find use in methods ofdelivering active agents to a topical location of a subject, where thetopical location may be a skin surface location or a mucosal location.In delivering active agents to a topical location of a subject, deliverycompositions of the invention may deliver the active agent-particlecomplexes at least into an epidermal location that is beneath the skinsurface of a subject. As such, embodiments of the invention includemethods of delivering active agent loaded particles into the stratumcorneum of a subject, where the methods may result in delivery of thecomplexes into the deep stratum corneum and/or dermis of a subject. By“into the stratum corneum” is mean that the complexes are delivered to aregion that is at least 1 cell layer below the skin surface. By “deepstratum corneum” is meant a region that is 2 or more cell layers belowthe skin surface, such as 5 or more cell layers below the skin surface,including 10 or more cell layers below the skin surface. In someinstances, the complexes are delivered to region of the stratum corneumthat is 2 μm or more such as 5 μm or more and including 15 μm or morebelow the surface of the skin.

Embodiments of the invention include methods of delivering active agentloaded particles into the stratum corneum of a subject, where themethods may result in delivery of the complexes into the dermis of asubject. By “into the dermis” is meant that the complexes are deliveredto a region that is at least 20 cell layers below the skin surface.

Upon reaching their target dermal location, in some instances the activeagent bound particles begin to release their active agent “payload”.Release of the active agent from the particles may occur according to anumber of different mechanisms. For example, the environment of the skinmay reverse any binding interaction of the agent to the particle. Inaddition to this mechanism or alternatively to it, the environment ofthe skin may break down the calcium phosphate particles (e.g., viadissolution caused by pH gradient of the skin), such that the uniform,rigid, spherical, nanoporous particles dissolve under acidic conditions,e.g., conditions of pH 5 or lower, such as 4.5 or lower, including 4.3or lower, such as the physiological acidic conditions of the stratumcorneum. The time required for dissolution of particles in the stratumcorneum may vary, and in certain embodiments ranges from 1 minute to 72hours, such as 10 minutes to 24 hours and including 30 minutes to 12hours, over which time period active agent is released from the activeagent bound particles. Aspects of the invention include release of allactive agent.

Methods of the invention therefore result in delivery of an active agentat least into the stratum corneum of a subject. In some embodiments, theactive agent remains in the stratum corneum to exert its desiredactivity. In yet other embodiments, the active agent may exert itsdesired activity at one or more other target locations of the body.Additional target locations interest include additional epidermalregions, such as but not limited to the stratum lucidum, stratumgranulosum, stratum spinusom, stratum basale and dermis. In certainembodiments, the active agent is delivered to a region of the dermis. Incertain embodiments, the active agent is delivered to a region below thedermis, e.g., into sub-cutaneous tissues.

In some instances the active agent may be systemically delivered to thesubject. When the active agent is systemically delivered to the subject,therapeutic plasma levels of active agent are achieved. Therapeuticplasma levels of active agent may vary depending on the particularactive agent and condition being treated. In certain embodiments,therapeutic active levels that are achieved range from 0.1 pg to 100 μg,such as 1 pg to 20 μg, such as 1 ng to 1 μg and including 10 ng to 100ng.

In practicing methods of the invention, a delivery composition isapplied to a topical region of a subject and maintained at the topicalregion in a manner sufficient to result in the desired delivery of theactive agent to the subject, as described above. The topical region is,in certain embodiments, a keratinized skin region. The keratinized skinregion, including hair follicles, sweat glands and sebaceous glands, maybe present at a variety of intact or damaged skin locations, wherelocations of interest include, but are not limited to: limbs, arms,hands, legs, feet; torso, e.g., chest, back, stomach; head, e.g., neck,face; etc. In certain embodiments, the region will be a head region,such as a facial region, e.g., forehead, occipital region, around themouth, etc. The topical region to which the composition is applied mayvary with respect to area, ranging in certain embodiments from 1 mm² to300 cm² or more, such as from 1 to 50 cm², and including from 3 to 10cm².

In practicing the subject methods, a subject may be administered asingle dose or two or more doses over a given period of time. Forexample, over a given treatment period of one month, 1 or more doses,such as 2 or more doses, 3 or more doses, 4 or more doses, 5 or moredoses, etc., may be applied to a topical location of the subject, wherethe doses may be applied weekly or daily or even multiple times per day.

Delivery of active agent complexes in accordance with the presentinvention may impart one or more advantages as compared to a controlwhich the active agent is not delivered as a complex with calciumphosphate particles. For example, in some instances the active agent isstabilized in the calcium phosphate complexes, such that its activity ispreserved. In some instances, the complexing the active agent withcalcium phosphate particle in complexes according to embodiments of theinvention provides for delivery of the active agent to locations inwhich it would not normally be delivered, e.g., delivery into thestratum corneum where delivery would be limited to the skin surface ifthat agent were not present in a calcium phosphate particle complex. Insome instances, methods of the invention result in enhanced penetrationof the active agent as compared to a suitable control. A suitablecontrol may be a delivery composition that includes the same activeagent and delivery vehicle components, but lacks the uniform, rigid,spherical, nanoporous calcium phosphate particles. In some instances,penetration is enhanced as compared to such a control by a factor of2-fold or more, such as 5-fold or more, including 10-fold or more. Inyet other embodiments, the complexes serve as a controlled release depotof active agent from the stratum corneum, thereby providing desiredextended release and delivery profiles for an active agent.

While the active agent-calcium phosphate complexes have been describedherein primarily in terms of their use for dermal delivery applications,in some instances they are employed for other applications. For example,the active agent-calcium phosphate complexes of the invention find usein some instances in non-dermal delivery of active agents to a subject.Examples of non-dermal delivery formulations include, but are notlimited to: capsules, tablets, pills, pellets, lozenges, powders,granules, syrups, elixirs, solutions, suspensions, emulsions,suppositories, or sustained-release formulations thereof, or any otherform suitable for administration to a mammal. In some instances, thepharmaceutical compositions are formulated for administration inaccordance with routine procedures as a pharmaceutical compositionadapted for oral or intravenous administration to humans. Examples ofsuitable pharmaceutical vehicles and methods for formulation thereof aredescribed in Remington: The Science and Practice of Pharmacy, Alfonso R.Gennaro ed., Mack Publishing Co. Easton, Pa., 19th ed., 1995, Chapters86, 87, 88, 91, and 92, incorporated herein by reference.

In certain embodiments, methods of delivering calcium into at least intothe stratum corneum are provided. In these methods, intact calciumphosphate particles of the invention are delivered into at least thestratum corneum, e.g., as described above. By “intact” is meant that theparticles are full integrity, undamaged particles. As such, they willnot be the same as particles that have been contacted with a chelatingagent, such as EDTA, where the chelating agent compromises the structureof the particles, e.g., by action of the chelating action with thecalcium ions. In these embodiments, the calcium phosphate particles maybe free of any bound active agent, e.g., they are calcium phosphateparticles that are not associated with an active agent. In theseembodiments, the delivery vehicle component may be free of any chelatingagent, e.g., EDTA. These methods find use in delivery calcium into atleast the stratum according for any convenient purpose, and may beperformed on subjects that are desirous of delivering calcium into atleast the stratum corneum. Any of the delivery vehicles described abovemay be employed, where those vehicles that are free of a chelating agentare of interest.

The subject methods and compositions may be used in a variety ofdifferent kinds of animals, where the animals are typically “mammals” or“mammalian,” where these terms are used broadly to describe organismswhich are within the class mammalia, including the orders carnivore(e.g., dogs and cats), rodentia (e.g., mice, guinea pigs, and rats),lagomorpha (e.g., rabbits) and primates (e.g., humans, chimpanzees, andmonkeys). In certain embodiments, the subjects or patients are humans.

The following examples are offered by way of illustration and not by wayof limitation.

Experimental I. Production and Characterization of Uniform, Rigid,Spherical, Nanoporous Calcium Phosphate Particles A. Production

A calcium phosphate nano-crystal slurry was prepared by dropwiseaddition of an aqueous phosphate complex solution into an aqueouscalcium complex solution or suspension under controlled conditions oftemperature, pH, pressure, gas, stirring velocity, reagentconcentration, addition rate and aging time. The slurry was spray driedto form a spherical porous powder by using a pressure nozzle type spraydryer with an air-liquid fluid nozzle. The dried powder was sintered attemperature ranging 300 to 900° C. for a period of time ranging 1 to 24hours with gas or electric furnace or kiln.

B. Characterization

FIGS. 1A and 1B show the porous structure of the resultant 2 micronuniform, rigid, spherical, nanoporous calcium phosphate particles(produced as described above) using SEM (A) 10,000×, (B) 50,000×. FIGS.2A and 2B show the outside and inside structure of the 2 micron uniform,rigid, spherical, nanoporous calcium phosphate particles (produced in asdescribed above) using using both SEM (A) and TEM (B) (15000×). Thelarge (25-50 m²/g) internal and external surface areas are substantial,allowing for high capacity binding with active agents. FIG. 3 shows theparticle size distribution of the particles, as determined by CoulterMulti-sizer 3 particle counter and confirmed by scanning electronmicroscopy. The average particle size was 2 μm.

C. Safety of Calcium Phosphate Particles

Cytotoxicity US-FDA 21 CFR Part 58 Non Toxic Mutagenicity Ames Test NonMutagenic Skin Sensitization RIPT (Human) Non Irritating

II. Preparation of Active Agent-Calcium Phosphate Particle Complexes

A. General Binding Guidelines Including Pre-treatment of certain ActiveAgents

Calcium phosphate particles bind a broad range of biomolecules and insome instances stabilizes them. The binding with calcium phosphateparticles is based on ionic interaction. The functional groups ofcalcium phosphate particles consist of positively charged calcium ions(Ca⁺⁺), and negatively charged phosphate ion (PO₄ ⁻³). This means theamount of the anionized carboxyl group of the biomaterials will bereduced under acidic conditions. Thus the binding between anionizedcarboxyl group of the biomolecules and calcium ion of calcium phosphateparticles will be weakened. The situation is reversed for theinteraction between the cationized amino group of the biomolecule andphosphate functional group of calcium phosphate particles in basicconditions.

The pH and ionic strength directly influence the binding between calciumphosphate particles and biomaterials. Calcium phosphate particles havethe ability to bind biomaterials with a broad range of molecular weights(e.g., 200 to 10,000,000) and isoelectric points (e.g., 2.0 to 12).

In addition to pH and ionic strength, the molecular weight, shape, andthe orientation of biomaterials also influence the binding to calciumphosphate particles. For example, BSA with relatively low molecularweight bind at 90 mg/g and DNA at a relatively large molecular weightbinds to calcium phosphate particles at the rate of 1 mg/g. The bindingcapacity of large biomolecules such as DNA will be determined by theouter surface area of calcium phosphate particles. To summarize, themain parameters influencing the binding between calcium phosphateparticles and biomolecules are pH, ionic strength, the stereochemicaleffect, and molecular weight.

B. Specific Active Agent-Calcium Phosphate Particle Complexes 1.Influence of pH on Binding

a. Salicylic Acid

Materials:

-   Calcium phosphate particles-   Salicylic acid, Fisher Scientific, Part No. A277-500

Methods:

-   i. 23.2 mg of Salicylic acid was dissolved in 1 ml of ethanol.-   ii. 4 g of calcium phosphate particles were suspended in 39.8 ml of    water and the pH was adjusted close to the targeted pH with HCl.-   iii. 0.2 ml of Salicylic acid solution (23.2 mg/ml) was mixed in the    suspension of calcium phosphate particles.-   iv. The pH was adjusted to each of the target pH values (11.36,    8.34, 7.47, 7.07, 5.99) with HCl.-   v. At each pH, a 4.8 ml sample was taken from the suspension. All    the samples were centrifuged separately at 2000×g for 10 min.-   vi. The absorbance of supernatant was measured at 297 nm (the    detection wavelength for Salicylic acid) by UV spectrophotometer.-   vii. The control was carried out by the same procedures without    calcium phosphate particles.

Results:

The results are shown in the table below and demonstrate that Salicylicacid binds to calcium phosphate particles in a pH dependent mannerdriven by the pKa of Salicylic acid.

Salicylic acid pH Bound (ug/g) 11.36 1141.14 8.34 660.4 7.47 97.5 7.0723.8 5.99 0b. Polyphenol Complex (PPC) to Calcium Phosphate Particles

Materials:

-   Calcium phosphate particles-   Polyphenol complex (PPC)

Methods:

-   i. 33.98 mg of PPC was dissolved in 6.8 ml of water.-   ii. 2 g of calcium phosphate particles were suspended in 19.9 ml of    water and the pH was adjusted to pH 9.62 with HCl.-   iii. 0.1 ml of PPC solution (5 mg/ml) was mixed in the suspension of    calcium phosphate particles; a 2 ml sample was taken.-   iv. The pH was adjusted to each of the target pH values (8.58, 8.07,    7.49, 7.21, 6.75, 6.08) with HCl.-   v. At each pH, a 2 ml sample was taken from the suspension. All the    samples were centrifuged separately at 2000×g for 10 min.-   vi. The absorbance of each supernatant was measured at 280 nm (the    detection wavelength for PPC) by UV spectrophotometer.-   vii. The control was carried out by the same procedures without    calcium phosphate particles.

Results:

Binding capacity of calcium phosphate particles is summarized in thetable below. The results demonstrate that PPC binding can be carried outat any pH, and is therefore pH independent.

pH PPC bound (μg/g) 9.62 242.5 8.58 242.5 8.07 235.0 7.49 215.0 7.21220.0 6.75 212.5 6.08 197.52. Binding Examples of Protein Actives with Different Molecular Weights(MW) and Isoelectric Points (pI)a. Bovine Serum Albumin (BSA) (MW: 66 KD, pI: 4.7)

Materials:

-   Calcium phosphate particles-   BSA, lypholized powder, Fisher Scientific, Catalog No. BP-671-10

Methods:

-   i. 0.5 g of calcium phosphate particles was suspended in 1 ml of    water, pH was adjusted with HCl to approximately 7. The suspension    was mixed for 10 min.-   ii. BSA was dissolved in water and gently mixed to prepare a 20    mg/ml solution. 4 ml of the BSA solution was added into each    suspension of calcium phosphate particles, and mixed for 30 min. The    final pH of the suspension was determined.-   iii. The suspension was centrifuged at 2000×g for 5 min. The    supernatant was transferred to new tubes and centrifuged again at    2000×g for 5 min.-   iv. A size-exclusion HPLC method was developed to quantify BSA in    the supernatants of the binding suspensions. The separation was    performed using a Phenomenex BioSep™-SEC-S3000 column (7.8×300 mm, 5    μm) in the Shimadzu 10AS system. The mobile phase was 100% 50 mM    phosphate buffer (Na⁺, pH 6.8), and eluted at a rate of 1.4 ml/min.    The eluent was monitored at 280 nm. BSA was observed as a major peak    with retention time at about 6.8 min. The quantification of BSA was    achieved by external standard calibration.-   v. The control was carried out by the same procedure without calcium    phosphate particles.

Results:

BSA bound to calcium phosphate particles at 95.1 mg/g.

b. Lactoferrin (MW: 90KD, pI: 8.5)

Materials:

-   Calcium phosphate particles-   Lactoferrin from human milk, Sigma Aldrich, Catalog No. 0520-100MG

Methods:

-   i. Lactoferrin solution was prepared in water at 4.98 mg/ml.-   ii. 0.3 g of Calcium phosphate particles was suspended in 1.2 ml of    water and mixed for 5 min.-   iii. 1.8 ml of Lactoferrin solution at 4.98 mg/ml was added to    achieve the final concentrations at 3.0 mg/ml, and the final volume    was 3 ml.-   iv. The suspension was mixed for 30 min, and centrifuged at 5000×g    for 10 min in a bench top centrifuge.-   v. The absorbance of supernatant was measured at 280 nm (the    detection wavelength for Lactoferrin) by UV spectrophotometer.-   vi. The control was carried out by the same procedures without    calcium phosphate particles.-   vii. Lactoferrin bound was calculated by subtracting the amount of    the Lactoferrin detected in the supernatant from the total initial    amount in the binding suspension.

Results:

29.63 mg/g Lactoferrin was determined bound to calcium phosphateparticles at a binding concentration of 2.99 mg/ml.

c. Lysozyme (MW: 14 KD, pI: 10.7)

Materials:

-   Calcium phosphate particles-   Lysozyme, MP biomedicals LLC. Product No. ICN10083405-   Phosphoric acid, Fisher Scientific, Product No. A260500

Methods:

-   i. 364.7 mg of Lysozyme as dissolved in 18.01 ml of water (20.25    mg/ml).-   ii. 0.8 g of calcium phosphate particles were mixed with 4 ml of    water, and the pH of the suspension was adjusted to neutral with    diluted phosphoric acid.-   iii. 4 ml of Lysozyme solution (20.25 mg/ml) was added to the    suspension of calcium phosphate particles to achieve a final    concentration of 10.124 mg/ml, and a final volume of 8 ml. The final    pH was measured.-   iv. The suspension was mixed for 30 min, and centrifuged at 2000×g    for 10 min in a bench top centrifuge.-   v. The absorbance of supernatant was measured at 280 nm (the    detection wavelength for Lysozyme) by UV spectrophotometer.-   vi. The control was carried out by the same procedures without    calcium phosphate particles.

Results:

Lysozyme was determined bound to calcium phosphate particles at 6.8 mg/gat pH 6.83.

3. Influence of Solvent on Binding

Adapalene

Materials:

-   Calcium phosphate particles-   Adapalene, Sekhsaria Chemicals Limited, India-   DMSO, Fisher Scientific, Product No. D159-4-   Ethanol, Fisher Scientific, Product No. AC61511-0010

Methods:

-   i. Adapalene was prepared as saturated solutions in ethanol and DMSO    (0.087 mg/ml in ethanol, 20.65 mg/ml in DMSO)-   ii. 0.5 g of calcium phosphate particles were mixed with 5 ml of the    Adapalene saturated solution in ethanol or DMSO.-   Iii. The suspensions were mixed for 30 min, and centrifuged at    2000×g for 10 min in a bench top centrifuge.-   iv. The absorbance of supernatants were measured at 319 nm (the    detection wavelength for Adapalene) by UV spectrophotometer.-   v. The controls were carried out by the same procedures without    calcium phosphate particles.

Results:

Binding capacity of Adapalene to calcium phosphate particles in ethanolis 0.78 mg/g. Binding capacity of Adapalene to calcium phosphateparticles in DMSO is 12.55 mg/g.

C. Example of Pretreatment of Bioactives

-   Argireline to calcium phosphate particles was pretreated with sodium    lauryl sulfate

Materials:

-   Calcium phosphate particles-   Argireline, (Acetyl Hexapeptide-8), Lipotec S.A.-   Sodium lauryl sulfate (SLS), Colonial Chemical, Inc.

Methods:

-   i. Argireline was dissolved in water as a 10 mg/ml solution.-   ii. Calcium phosphate particles were mixed with 10 ml of 0.1% SLS    for 5 min. The suspension was centrifuged at 2000×g for 10 min in    the bench top centrifuge, and the supernatant was removed. The    pellet was re-suspended in 20 ml water, centrifuged at 2000×g for 10    min, and the washing supernatant was discarded. The washing step was    repeated twice. The water contained in the pellet was determined.    The final pellet was used in the binding study.-   iii. Calcium phosphate particles (SLS treated or no SLS treatment)    were mixed with water. HCl (or NaOH) was added to adjust the pH of    Calcium phosphate particles suspension to targeted pH including    neutral and pH ˜10.-   iv. Argireline stock at 10 mg/ml was added to each binding    suspensions to be a final concentration of 0.5 mg/ml. After the    suspension was mixed for 30 min, the final pH was measured.-   v. The binding suspensions were centrifuged at 2000×g for 10 min in    the bench top centrifuge. The supernatants were analyzed with    Refractive Index detector connected to the Shimadzu HPLC 20A system    to quantify the free Argireline in the solution. A size-exclusion    HPLC method was developed to quantify Argireline in the supernatants    of the binding mixtures. The separation was achieved using a    Phenomenex BioSep™-SEC-S3000 column (7.8×300 mm, 5 μm) in the    Shimadzu 20A system. The mobile phase was 100% water, and eluted at    a rate of 1 ml/min. The eluent was monitored at 205 nm or by a    refractive index detector (Shimadzu, Model No. RID-10A). Argireline    was observed as a major peak in the chromatogram with retention time    of about 14 min. The quantification of Argireline was achieved by    external standard calibration.-   vi. The Argireline bound was then calculated by subtracting the    amount of Argireline detected in the supernatant from the total    initial amount in the binding suspension.

Results:

Calcium phosphate Argireline bound particles pH (mg/g) SLS treated 7.851.19 SLS treated 10.10 2.15 No pretreatment 7.39 0.09 No pretreatment10.46 0.05

D. Visualization of Active Agent Bound to Calcium Phosphate Particles

0.1 g of calcium phosphate particles was added to 1 ml 0.9% Rhodamine Bin water, and the resultant suspension was spun and the supernatant wasremoved. The resultant Rhodamine B-calcium phosphate particles weredried at 58° C. for 24 hours. The resultant powder was resuspended inCaprylic/Capric Triglyceride and imaged via microscopy. The resultantimage is shown in FIG. 4A. FIG. 4B shows calcium phosphate particleswithout any Rhodamine B.

E. Additional Active Agent-Calcium Phosphate Complexes

Using the protocols illustrated above, active agent-calcium phosphatecomplexes were produced as summarized in the following tables. In thefollowing tables, the specific solvent systems are examples of solventsystems that may be used.

Classes Active Name Solvent 534-1. Azo Compounds Sulfasalazine Ethanol534-1. Diazo Compounds Azaserine Water 536-2. Carbohydrate derivativesDextran sulfate sodium salt Water MW 20000 536-2. Carbohydratederivatives Hyaluronic Acid, Low MW Water 536-2. Carbohydratederivatives Hyaluronic Acid, High MW Water 536-3. Glycosides Doxorubicinhydrochloride Water (Adriamycin) 536-4. Oxygen-containing hetero Uridine5′-diphosphoglucose 10 mM Bis-Tris ring disodium salt Buffer 536-5.Flavon sugar compounds Riboflavin 5′-monophosphate 10 mM Bis-Tris Buffer536-5. Flavon sugar compounds Rutin hydrate Ethanol 540-6. Steroidalhetero Sodium Cholesteryl Sulfate Ethanol compounds 540-7. AzaporphyrinsCyanocobalamin Acetonitrile 540-8. Four-membered lactam Cefaclor Waterwith avicinyl halogen 540-9. Nitrogen hetero rings of GYKI 52466hydrochloride Water more than six members (can include multipleheteroatoms) 544-10. Hetero ring is six- Thiamine Pyrophosphate Watermembered having two or more ring heteroatoms of which at least one isnitrogen 544-11. Six-membered hetero Acesulfame K Acetonitrile ringconsists of oxygen, sulfur, nitrogen and carbon 544-12. Six-memberedhetero Methylene Blue Acetonitrile ring consists of sulfur, nitrogen andcarbon 544-13. Six-membered hetero Furaltadone Dichloromethane ringconsists of oxygen, nitrogen and carbon 544-14. Six-membered heteroCiclopirox Olamine Water ring consists of nitrogen and carbon 544-15.Hetero ring is six- Nicotine Water membered consisting of one nitrogenand five carbons 548-16. Hetero ring is five- Imiquimod Ethanol memberedhaving two or more ring hetero atoms of which at least one is nitrogen548-18. Hetero ring is three- (1R)-(−)-(10- DMSO membered having two ormore Camphorsulfonyl)oxaziridine ring hetero atoms of which at least oneis nitrogen 549-19. Sulfur containing Amoxicillin Acetonitrile heteroring (e.g., thiiranes, etc.) See above. 549-20. Oxygen containing(−)Scopolamine methyl Water hetero ring (e.g., oxirane, nitrate etc.)See above 549-20. Oxygen containing Calcein Water hetero ring (e.g.,oxirane, etc.) See above 552-21. Azides AZT (Azidothymidine or DMSOZidovudine) 552-22. Triphenylmethanes o-Cresolphthalein Complexone 10 mMBis-Tris Buffer 552-23. Tetracyclo naphthacene Minocycline Hydrochloride(HCl), 10 mM Bis-Tris configured ring system having at Buffer least onedouble bond between ring members 552-23. Tetracyclo naphthaceneChlortetracycline HCl Water configured ring system having at least onedouble bond between ring members 552-24. Quinolines, HydrocarbonDifloxacin HCl Water 552-25. Steroids CHAPS (3-[(3- AcetonitrileCholamidopropyl)dimethylammonio]- 1-propanesulfonate) 554-26. Fattycompounds having Lecithin (L-α Phosphatidylcholine) Octanol an acidmoiety which contains the carboxyl of a carboxylic acid, salt, ester, oramide group bonded directly to one end of an acyclic chain of at leastseven uninterrupted carbons. 556-27. Heavy metal, aluminum, CarboplatinAcetonitrile or silicon organic compounds. 558-28. Thioimidate estersSinigrin Hydrate Water 558-29. Imidate esters Ethyl BenzimidateHydrochloride DMSO 558-30. Thiocyanate esters Allyl isothiocyanateHexane 558-31. Sulfate esters β-Estradiol 3-sulfate sodium salt DMSO558-32. Sulfonate esters Alizarin Red S Water 558-33. Phosphorus estersAlendronic Acid Acetonitrile-1M (phosphonate, phosphonic acid) NaOH558-35. Nitrate esters or Norcandil DMSO chalcogen analogues 560-36.Carboxylic acid esters Methyl Salicylate Ethanol 560-37. Sulfonic acids,salts, Chondroitin sulfate A sodium salt Water halides 560-38.Sulfohydroxamate esters Sulfadimethoxine Isopropyl Alcohol or chalcogenanalogues 560-39. Perhydroxamate esters 6-AminonicotinamideAcetonitrile-1M or chalcogen analogues HCl 562-40. Organic acidsSalicylic acid Acetonitrile 562-40. Organic acids L-Arginine Water562-40. Organic acids L-Histidine Acetonitrile-1M HCl 562-40. Organicacids DPHP (Dipalmitoyl Hydroxyproline) Ethanol 562-40. Organic acidsAdapalene DMSO 562-40. Organic acids Ca PCA (Calcidone) Water 562-41.Acid halides, acid Poly[(isobutylene-alt-maleic acid, Acetonitrileanhydrides ammonium salt)-co-(isobutylene-alt- maleic anhydride)],average Mw ~60,000 562-42. Selenium or Tellurium Seleno-DL-cystineIPA-1M HCl; compounds Acetic Acid-1M HCl 564-43. Ureas Allantoin DMSO564-44. Sulfonamides, sulfamides Sumatriptan Succinate Acetonitrile564-45. Nitro-containing 2-Nitrophenyl β-D-glucopyranoside IsopropylAlcohol compounds 564-46. Carboxamides Z-L-Asparagine Acetonitrile-Water 564-47. Oxyamines Methoxyamine hydrochloride Acetonitrile 568-48.Boron, Phosphorus, Resveratrol Ethanol Sulfur, or Oxygen compounds568-48. Boron, Phosphorus, ATP (Adenosine Triphophate) Water Sulfur, orOxygen compounds 568-48. Boron, Phosphorus, ADP (Adenosine Diphosphate)Water Sulfur, or Oxygen compounds 568-48. Boron, Phosphorus, AMP(Adenosine Monophosphate) Water Sulfur, or Oxygen compounds 568-48.Boron, Phosphorus, Zoledronic acid Acetonitrile-1M Sulfur, or Oxygencompounds NaOH 570-49. Halogen containing Diclofenac Acetonitrileorganic compounds 424-50. Lymphokines Gamma Globulins from bovine bloodWater 424-51. Enzyme or coenzyme Thrombin Topical (Recombinant) Water424-51. Enzyme or coenzyme Superoxide dismutase Water 424-52. Extract,body fluid, or Catalase Water cellular material of undeterminedconstitution derived from animal is active ingredient 424-52. Extract,body fluid, or Goat IgG Water cellular material of undeterminedconstitution derived from animal is active ingredient 424-53. Inorganicactive Bacitracin zinc salt Acetonitrile-1M ingredient containingcomposition, HCl e.g. metal 424-53. Inorganic active Copperphthalocyanine Water ingredient containing composition, e.g. metal424-54. Extract or material Streptomycin sulfate salt Acetonitrilecontaining or obtained from a multicellular fungus as active ingredient435-55. Enzyme (e.g., ligases, Sulfatase Water etc.), proenzyme; 435-55.Enzyme (e.g., ligases, Phosphatase Water etc.), proenzyme; 435-55.Enzyme (e.g., ligases, Lysozyme Water etc.), proenzyme; 435-56. Virus orbacteriophage, Bacteriophage CE6 Water; except for viral vector or λphage for delivery of T7 RNA 20 mM Na bacteriophage vector; polymerasePhosphate composition thereof; Buffer 435-57. Micro-organism Ampicillinsodium Water 435-57. Micro-organism Gentamicin sulfate from WaterMicromonospora purpurea 530-58. Peptides of 3 to 100 Lys-Lys-LysAcetonitrile amino acid residues 530-58. Peptides of 3 to 100 ArgirelineWater amino acid residues 530-59. Peptides containingN-Acetylmuramyl-L-alanyl-D- Ethanol unnatural amino acid residues orisoglutamine hydrate (Muramyl derivatives Dipeptide) 530-60. Peptidescontaining non- Cyclosporin A Hexane linear or heterogeneous backboneelements 530-61. Peptide-like structures Boc-Gly-Lys-Arg-7-amido-4-Acetonitrile containing terminal methylcoumarin hydrochloridefunctionalization(s) 530-62. Proteins, i.e., more than Lactoferrin(human precursor Water 100 amino acid residues reduced) 530-62.Proteins, i.e., more than BSA Water 100 amino acid residues 530-62.Proteins, i.e., more than Ovalbumin Water 100 amino acid residues530-62. Proteins, i.e., more than Phycoerythrin Water 100 amino acidresidues 426. Food or edible material: Fermented soybean extract Water;processes, compositions, and Glycerin products 426. Food or ediblematerial: Aloe Vera (Botanivera 1-200C) Water processes, compositions,and products 426-63. Product for promoting the Ascorbic Acid (Vitamin C)Water effect of an alimentary canal microorganism Product with addedvitamin or derivative thereof for fortification 426-63. Product forpromoting the Sodium Ascorbyl Phosphate Water effect of an alimentarycanal (Stay-C) microorganism Product with added vitamin or derivativethereof for fortification 426-63. Product for promoting the PotassiumAscorbyl Tocopheryl Water effect of an alimentary canal Phosphate(Sepivital) microorganism Product with added vitamin or derivativethereof for fortification 426-63. Product for promoting the DisodiumAscorbyl Sulfate (VC-SS) Water; effect of an alimentary canalAcetonitrile microorganism Product with added vitamin or derivativethereof for fortification 426-63. Product for promoting the UbiquinolHexane effect of an alimentary canal microorganism Product with addedvitamin or derivative thereof for fortification 426-64. PredominantlyUrsolic Acid DMSO hydrocarbon compounds containing cyclic carbon rings;three-, four-, five-, six- or more membered rings.

Loading Optimum Structural No Actives Solvent pH classes 1 Ascorbic acidWater >8.12 Sugar acids 2 Salicylic Acid Ethanol/ 4.03-7.90 Aromaticwater acids 3 Hyaluronic acid Water 7 Polysaccharide Sodium salt 4Hyaluronic acid Water 7 Polysaccharide Sodium salt 5 Argireline Water6.7-7.8 Peptides Water 7.8 Water 6.4 6 Fermented Soybean Glycerin 7Proteins Extract 7 Sepivital (dl-a- Water 7 Vitamin tocopheryl 2 Lderivatives ascorbyl phosphate) 8 Sepilift DPHP Ethanol Amino acid(dipalmitoyl derivatives hydroxyproline) 9 Resveratrol Ethanol/ 10Polyphenol water 10 Bovine serum albumin Water 7 Proteins 11Chlortetracycline Water   pH 7-8.5 Tetracycline antibiotic, 12Ciclopirox Olamine Water 7 Alkaloids Water pH 8.5 Water pH 8.5 Water pH9.1 Water pH 8.9 13 Adapalene Ethanol 11 Retinoids Ethanol 7 DMSO 7 14Imiquimod Ethanol 11 Alkaloids Ethanol 70.8 15 Adriacin Water 16 Alphalipoic acid Water 17 Green tea polyphenol Water Polyphenol 18 MatrixylAcetate Water Peptide (PalmitoylPentapeptide, PAL-Lys-Thr-Thr-Lys-Ser)19 Oleic acid No solvent Fatty acid 20 Oleyl oleate No solvent Fattyacid 21 Dextran Water polysaccharide 22 Ascorbic acid Water Vitamin Cglucoside derivative

III. Release of Active Agent A. pH Dependent Release 1. Lysozyme

In the stratum corneum, the pH ranges from approximately 4.3 to 5.0 withpH decreasing with stratum corneum depth. To study calcium phosphateparticle release active agent at conditions analogous to skin, twolysozyme calcium phosphate complexes were exposed to buffers of ph 4.8(0.5 M sodium acetate) and 7.0 (10 mM Bis Tris) for 8 hours. The buffersflowed through the samples at a rate of 1 mL/hr and were collectedhourly and analyzed for lysozyme release by monitoring the peak at 280nm by UV spectrometer. An equivalent mass of lysozyme Calcium phosphateparticles complex was vortexed in pH 4.8 buffer to estimate the totalavailable lysozyme in the sample. At pH 4.8, lysozyme was observed toquickly release from the calcium phosphate particles with most of thelysozyme releasing within the first hour. In contrast, at pH 7, nolysozyme is observed released from calcium phosphate particles over 8hours.

B. Release Via Hydroxysome Degradation

1. Calcium phosphate particles degrade at pH 4.8

50 mg of calcium phosphate particles were incubated in 2 mL at two pHs:4.8 (0.5 mM sodium acetate) and 7.1 (0.1 mM Bis Tris) and the solutionsplace on a rotator for 96 hours at room temperature. The samples werecentrifuged and the pellet dried and weighed. The percent weight loss ofcalcium phosphate particles at pH 4.8 and 7.1 was 12% and 3%,respectively. The buffering capacity of this closed system limited thecomplete dissolution of the calcium phosphate particles. Dissolution wasthen studied in a flow-through system with the same buffer whereby thesolution was slowly (5 mL/hr) flowed through the sample (50 mg) withgentle agitation of the sample followed by collection and drying of thecalcium phosphate particle pellet. After 72 hours, the percent weightloss of calcium phosphate particle pellet at pH 4.8 and 7.1 was 31% and3%, respectively. These results demonstration that calcium phosphateparticles dissolve at low pH and this dissolution is a function of thepH and buffering capacity of the solution.

C. Active Agent is Reversibly Bound to Calcium Phosphate Particles andRelease Does Not Alter Activity of Active Agent 1. BSA Calcium PhosphateComplex

BSA-calcium phosphate complexes were prepared as above. The resultantcomplexes were washed with water and then treated with 0.2 M sodiumphosphate to release any bound BSA.

Materials:

-   Calcium phosphate particles-   BSA, lypholized powder, Fisher Scientific, Catalog No. BP-671-10

Methods:

-   i. 0.1170 g of BSA was dissolved in 11.7 ml of water as a solution    of 10 mg/ml.-   ii. 0.5 g of calcium phosphate particles were mixed with 5 ml of BSA    solution at 10 mg/ml. The suspension was mixed for 30 min and the    final pH was determined to be neutral.-   iii. The suspension was centrifuged at 2000×g for 10 min. The    supernatant was transferred to new tubes and centrifuged again at    2000×g for 10 min.-   iv. The final supernatant was analyzed with Shimadzu 10A HPLC system    to quantitate BSA and calculate the binding.-   v. The pellet from 5 ml of binding suspension was mixed with 0.8 ml    of water, and centrifuged at 2000×g for 10 min. The rinsed pellet    was mixed again with 0.8 ml of water and centrifuged at 2000×g for    10 min.-   vi. The final rinsed pellet was mixed with 2 ml of 500 mM sodium    phosphate buffer (pH 6.8) and 2.235 ml of water to release BSA in a    suspension with 200 mM sodium phosphate. The release suspension was    centrifuged at 2000×g for 10 min. The supernatant was analyzed with    Shimadzu 10A HPLC system to quantitate BSA.-   v. The BSA quantitation was achieved using a Phenomenex    BioSep™-SEC-S3000 column (7.8×300 mm, 5 μm) in the Shimadzu 10AS    system. The mobile phase was 100% 50 mM phosphate buffer (Na⁺, pH    6.8), and eluted at a rate of 1.4 ml/min. The eluent was monitored    at 280 nm. BSA was observed as a major peak with retention time at    about 6.8 min. The quantification of BSA was achieved by external    standard calibration.-   vi. The control was carried out by the same procedures without    calcium phosphate particles.

Results:

The released BSA was analyzed with HPLC and determined to be identicalto an unbound control (retention time at 6.85 min), demonstrating thatbinding and release to calcium phosphate particles did not impact BSAintegrity.

2. Sodium Tocopheryl Phosphate Calcium Phosphate Complex Materials:

-   Calcium phosphate particles-   Sodium Tocopheryl Phosphate (TPNa), Showa Denko KK-   Ethanol, Fisher Scientific, Product No. AC615090020

Methods:

-   i. 20 mg of TPNa was dissolved in 40 ml of water by gently mixing    (0.5 mg/ml).-   ii. 3 g of calcium phosphate particles was mixed with 30 ml of TPNa    solution at 0.5 mg/ml, and mixed for 30 min.-   iii. The binding suspension was centrifuged at 2000×g for 10 min.    The supernatant was transferred to new tube, and centrifuge again at    2000×g for 10 min to clarify. The final supernatant was analyzed    with an UV spectrophotometer at 286 nm to quantitate free TPNa and    calculate the binding. Approximately 100% of TPNa in the binding    suspension was attached to calcium phosphate particles.-   iv. The pellet of TPNa calcium phosphate complex was re-suspended in    60% ethanol to release the bound TPNa.-   v. The release suspension in 60% ethanol was centrifuged at 2000×g    for 10 min. The supernatant was analyzed with an UV    spectrophotometer at 286 nm to quantitate free TPNa released.

Results:

The released TPNa was analyzed with UV spectroscopy and determined to beidentical to an unbound control, demonstrating that binding and releaseto calcium phosphate particles did not impact TPNa integrity.

IV. Examples of Formulations 1. Calcium Phosphate Particle—RiboflavinMonophosphate Ointment Formulation

Trade Name INCI Name w/w % 1 Bee wax Bee wax 5.00 2 SonneNatural ™Glyceride oils 74.66 3 Protachem IPP Isopropyl Palmitate 15.00 4 CapmulMCM Glyceryl 3.33 Caprylate/Caprate 5 Purified Water Water 1.00 6calcium Hydroxyapatite 1.00 phosphate particles 7 Riboflavin Riboflavin0.0077 Monophosphate monophosphate

Procedure:

-   Step 1. In a beaker add Bee wax and Protachem IPP. Start to heat to    70° C.-75° C. until uniform. Cool to 50° C. Add SonneNatural™ and    Capmul MCM. Mix until uniform.-   Step 2. In a separate beaker add 1% water and Riboflavin    monophosphate. Mix at R.T until dissolved. Add calcium phosphate    particle (adjust pH with lactic acid to pH 7). Spin for 10 minutes    at 500 RPM.-   Step 3. Transfer Step 1 to Step 2 under room temperature. Mix until    uniform

B. Stability of Active Agent in Formulation 1. Vitamin C

a. Methods:

11.9 mg of ascorbic acid was dissolved in 30 ml of water. 2 g of 2 μmcalcium phosphate particles were suspended in 19.5 ml of water and thepH was adjusted to pH 7.14 with HCl. 0.5 ml of ascorbic acid solution(0.40 mg/ml) was mixed in the suspension of calcium phosphate particles,which reduced the pH to 7.05. The suspension (pH 7.05) of calciumphosphate particles including 10 μg/ml of ascorbic acid was incubated at50° C. for 0.5 to 5 hours. The heat-denaturation of ascorbic acid wasterminated by cooling the suspension in an ice bath for 15 minutes. Inorder to release bound ascorbic acid from calcium phosphate particles tomeasure the stabilization effect, the pH of each suspension was adjustedto pH 5 with HCl and centrifuged at 3,000 rpm for 10 minutes. Thesupernatant obtained was measured at 265 nm. As a control, a 10 μg/ml ofascorbic acid solution was also incubated and measured by the sameprocedures without calcium phosphate particles. The activity of ascorbicacid was calculated by the following formula:

The activity of ascorbic acid (%)=(A _(265nm) of the supernatant afterincubation/A _(265nm) of the ascorbic acid before incubation)×100

The same procedures were repeated with 10 μm calcium phosphateparticles.b. Results

The results are summarized in the table below. The activity of Ascorbicacid was reduced to 36% by incubation at 50° C. for 0.5 hrs; thisactivity was increased to 85% when ascorbic acid was attached to 2 μmcalcium phosphate particles during the same period. The ascorbic acidwithout calcium phosphate particles was totally denatured with noactivity after the incubation at 50° C. for 3 hrs. However, the ascorbicacid bound to 2 μm calcium phosphate particles had 48% activity at 50°C. for the same 3 hr period.

Time (hrs) Control (%) 2 um (%) 0 100 100 0.5 36 85 2 18 59 3 0 48 5 035C. In order to achieve stability of the active in the calcium phosphatecomplex in final formulation, it may be necessary to add additional freeactive to the solvent system depending on the solubility of the solventsystem used.

V. Delivery Studies

1. Delivery into Stratum Corneum in Human Skin

A suspension of calcium phosphate particles in water at a concentrationwas applied to a forearm of a living human by rubbing for 10 seconds(FIG. 5). Tape stripping of the first, second and third layers of thestratum corneum was then performed. The calcium phosphate particlespenetrated to the third layer of the stratum corneum.

2. Delivery into Lower Layers of Stratum Corneum in Mouse Skin

Calcium phosphate particles penetrate the stratum corneum and penetratefurther into the lower layers as the particle disintegrates into smallersubstituent parts. Calcium phosphate particles were no longer intactafter 7 hours and were no longer spherical, indicating loss of theintegrity of the particles.

a. Materials:

2 μm calcium phosphate particles were used.

b. Preparation:

The calcium phosphate particles were suspended in 70% Ethylene glycoland 30% Ethanol to make a 10% suspension. The resultant suspension wastopically applied on the skin surface of hairless mice (1×1 cm area).During the application, mice were given anesthetic. The firstapplication (0.2 ml) was administrated and left on, the secondapplication was administrated 4 hours later at the same place and forthe same amount. Seven hours from the first application the skin wasremoved, and treated by the Ca⁺⁺. Localization method followed by EMtechniques.

c. Results:

Prior to treatment with calcium phosphate particles, examination of theepidermis reveals only in the areas of Stratum granulosum, with nodetectable Ca⁺⁺ in stratum corneum (FIG. 6). After topical applicationof calcium phosphate particles as described above, calcium phosphateparticles can be seen in the stratum corneum (FIG. 7A), with thesmallest particles moving into deeper layers (FIG. 7B).

3. Active Agent Detection in the Stratum Corneum

a. Purpose of Study

The purpose of this study was to detect active agent (chlorotetracycline(CTC)) in the stratum corneum after the topical application of activeagent (CTC) attached to 2 μm uniform, rigid, spherical, nanoporouscalcium phosphate particles, as described in Example I, above.Chlorotetracycline (CTC from Sigma, part #C-4881) was selected becauseit allows the visualization of CTC by fluorescence and is thusdetectable by confocal microscopy within the stratum corneum.

b. Preparation

A CTC solution was made by dissolving 80 mg of CTC in 10 ml of water.The un-dissolved CTC was removed by centrifugation at 3,000 rpm for 10min. 200 mg of particles were mixed with 2 ml of the CTC solution andvortexed for 1 min. The free CTC was removed by 3 cycles of washing withwater followed by centrifugation for 10 min at 3,000 rpm. The resultantCTC bound particles were suspended in water at 1:10 dilution rate.

c. Topical Application

The suspension produced in 3.b above (approximately 0.2 ml) wastopically applied onto the skin of hairless mice (1×1 cm area). Duringthe application, mice were given anesthetic. After 7 hours their skinswere examined using confocal microscopy (at 510 nm wavelength emissionexcited at 380 nm wavelength).

d. Results

The resultant confocal microscopy image is shown in FIG. 8(magnification is 300× in FIG. 8). The representative image shown inFIG. 8 shows that CTC fluorescence (purple) penetrates the skin and ismainly localized in the stratum corneum.

4. Tape-Stripping Analysis of Topical Application of STAY-C50-CalicumPhosphate and Lysozyme-Calcium Phosphate Complexes Demonstrate DeliveryInto the Stratum Corneum

The purpose of this study was to detect the distribution of actives inthe stratum corneum following topical application of the active-calciumphosphate particles

-   Method: The distribution of STAY C50 within the stratum corneum was    assessed by serial tape stripping of the skin to which a STAY    C50-calcium phosphate particle formulation was applied. An    application site on the forearm of a human subject was marked and    200 μl of the formulation spread by spatula. The site was allowed to    dry for 10 minutes and then followed by ten pre-weighed tape    stripping using strips of (3 in²) applied to the application site.    STAY C50 was extracted from the tape strips by sonicating the    samples in water for 30 minutes and analyzing the samples by HPLC.    The tape strips showed delivery of STAY-C50 to the 10th layer of    stratum corneum (FIG. 9).-   Method: The distribution of lysozyme within the stratum corneum was    assessed by serial tape stripping of the skin to which a    lysozyme-calcium phosphate particle formulation was applied. An    application site on the forearm of a human subject was marked and    200 μl of the formulation spread by spatula. The site was allowed to    dry for 10 minutes and then followed by ten pre-weighed tape    stripping using strips of (3 in²) applied to the application site.    Lysozyme was extracted from the tape strips by sonicating the    samples in water for 30 minutes and analyzing the samples by HPLC.    Lysozyme was detected to a depth of 6 tape strips (FIG. 10).

5. Controlled Slow Release of Active Agents by Franz Cell

-   Purpose: The purpose of this study was to detect riboflavin from    riboflavin attached to calcium phosphate particles.

Methods:

-   Circular 6 cm² discs of full-thickness pig skin were cut from a    larger abdominal specimen. Fat was removed from the dermis-side by    scissors and the skin was stored at −20° C. until use. The skin was    affixed between the two compartments of the glass diffusion cell    (Laboratory Glass Apparatus, Model #LG-1084-LPCT). This allowed an    exposed skin area of 5 cm² over a receptor compartment volume of    4.5 ml. The diffusion cell was maintained at 37° C.

Penetration Conditions

-   -   Topical application intervals were 8 and 16 hrs. The cells were        covered with parafilm and shielded from light by aluminum wrap.        A receptor fluid of phosphate buffered saline (PBS) was spun at        60 rpm.

Applied Formulations

-   -   Riboflavin monophosphate-calcium phosphate particles were        prepared to deliver 0.35-1.15 mg riboflavin (20-38%        suspensions).    -   Riboflavin monophosphate in PBS was prepared at 1.5 mg/mL to        deliver 0.3-0.45 mg riboflavin.    -   Formulations were applied by pipette in volumes of 50-100 μl.        Riboflavin-calcium phosphate complex applications were allowed        to air dry and then equal volume of 0.5 M sodium acetate buffer        was pipetted onto the application site. Controls consisted of        equivalent applications of riboflavin without calcium phosphate        particles.

Sample Collection

-   -   The skin surface, while still retained in the diffusion cell,        was washed twice with 1 mL PBS each time. The skin was removed        from diffusion cell and dried and analyzed.

Sample Analysis

-   -   Washes and Receptor fluid: The approximate volume of the fluids        was determined. The samples were spun at 10,000 rpm for 30        seconds and the calcium phosphate particles separated, dried and        weighed. The supernatant was then removed and analyzed by UV        spectrometer. If the UV absorbance was saturated (over >2.0),        the samples were diluted with water. The UV absorbance value at        370 nm was recorded.    -   Skin: The skin was minced and mixed with 5 ml of 10%        trichloracetic acid solution. The sample was sonicated with        ultrasonic apparatus at 50° C. for 1 hour. The sample was        centrifuged at 10,000 rpm for 10 minutes, the supernatant        removed and analyze by UV spectrometer. If the UV absorbance was        saturated (over >2.0), samples were diluted with water. The        maximum UV absorbance value at 370 nm was recorded.

Results

As detected by the appearance of riboflavin metabolite in the receptorfluid after 8 and 16 hours, riboflavin penetrated through skin from boththe aqueous solution and from calcium phosphate particle complex. Thecalcium phosphate particle complex caused a slower release of theactive. The results are shown graphically in FIG. 11.

Although the foregoing invention has been described in some detail byway of illustration and example for purposes of clarity ofunderstanding, it is readily apparent to those of ordinary skill in theart in light of the teachings of this invention that certain changes andmodifications may be made thereto without departing from the spirit orscope of the appended claims.

Accordingly, the preceding merely illustrates the principles of theinvention. It will be appreciated that those skilled in the art will beable to devise various arrangements which, although not explicitlydescribed or shown herein, embody the principles of the invention andare included within its spirit and scope. Furthermore, all examples andconditional language recited herein are principally intended to aid thereader in understanding the principles of the invention and the conceptscontributed by the inventors to furthering the art, and are to beconstrued as being without limitation to such specifically recitedexamples and conditions. Moreover, all statements herein recitingprinciples, aspects, and embodiments of the invention as well asspecific examples thereof, are intended to encompass both structural andfunctional equivalents thereof. Additionally, it is intended that suchequivalents include both currently known equivalents and equivalentsdeveloped in the future, i.e., any elements developed that perform thesame function, regardless of structure. The scope of the presentinvention, therefore, is not intended to be limited to the exemplaryembodiments shown and described herein. Rather, the scope and spirit ofpresent invention is embodied by the appended claims.

What is claimed is:
 1. A composition comprising uniform, rigid,spherical nanoporous calcium phosphate calcium phosphate particles,wherein the particles have an average particle diameter of 2 microns orless and are complexed with an active agent.
 2. The compositionaccording to claim 1, wherein the particles have a pore volume rangingfrom 30 to 85%.
 3. The composition according to claim 2, wherein theparticles have a pore size ranging from 2 nm to 100 nm.
 4. Thecomposition according to claim 1, wherein the particles are produced by:preparing a fluid composition of calcium phosphate crystals; drying thefluid composition in a manner sufficient to produce precursor particles;and subjecting the precursor particles to elevated temperature andpressure in a manner sufficient to produce uniform, rigid, sphericalnanoporous calcium phosphate calcium phosphate particles.
 5. Thecomposition according to claim 4, wherein the drying comprisesspray-drying.
 6. The composition according to claim 1, wherein thecomposition is a topical formulation.
 7. The composition according toclaim 1, wherein the amount of active agent complexed with the particlesranges from 0.01 to 300 mg active agent per gram of particles.
 8. Amethod of delivering an active agent to a subject, the methodcomprising: applying a composition comprising: uniform, rigid, sphericalnanoporous calcium phosphate calcium phosphate particles, wherein theparticles have an average particle diameter of 2 microns or less and arecomplexed with an active agent; to a topical region of the subject todeliver the active agent to the subject.
 9. The method according toclaim 8, wherein the particles have a pore volume ranging from 30 to85%.
 10. The method according to claim 9, wherein the particles have apore size ranging from 2 nm to 100 nm.
 11. The method according to claim8, wherein the amount of active agent complexed with the particlesranges from 0.01 to 300 mg active agent per gram of particles.
 12. Themethod according to claim 8, wherein the topical location is a mucosallocation.
 13. The method according to claim 8, wherein the topicallocation is a keratinized skin surface.
 14. The method according toclaim 13, wherein the method is a method of delivering the active agentat least into the stratum corneum of the subject.
 15. The methodaccording to claim 13, wherein the method is a method of delivering theactive agent into the deep stratum corneum of the subject.
 16. Themethod according to claim 13, wherein the method is a method ofdelivering the active agent through the stratum corneum.
 17. The methodaccording to claim 13, wherein the method is a method of delivering theactive agent into the dermis of the subject.
 18. The method according toclaim 13, wherein the method is a method of systemically delivering theactive agent to the subject.